Modulation of HIF1alpha and HIF2alpha expression

ABSTRACT

Compounds, compositions and methods are provided for modulating the expression of HIF1α and/or HIF2α. The compositions comprise oligonucleotides, targeted to nucleic acid encoding HIF1α and HIF2α. Methods of using these compounds for modulation of HIF1α and/or HIF2α expression and for diagnosis and treatment of disease associated with expression of HIF1α and/or HIF2α are provided.

FIELD OF THE INVENTION

[0001] The present invention provides compositions and methods formodulating the expression of HIF1α (HIF1α) and hypoxia-inducible factor2 alpha (HIF2α). In particular, this invention relates to compounds,particularly oligonucleotide compounds, which, in preferred embodiments,hybridize with nucleic acid molecules encoding HIF1α and HIF2α. Suchcompounds are shown herein to modulate the expression of HIF1α andHIF2α.

BACKGROUND OF THE INVENTION

[0002] Oxygen homeostasis is an essential cellular and systemicfunction; hypoxia leads to metabolic demise, but this must be balancedby the risk of oxidative damage to cellular lipids, nucleic acids, andproteins resulting from hyperoxia. As a result, cellular and systemicoxygen concentrations are tightly regulated via response pathways thataffect the activity and expression of a multitude of cellular proteins.This balance is disrupted in heart disease, cancer, cerebrovasculardisease, and chronic obstructive pulmonary disease (Semenza, Genes Dev.,2000, 14, 1983-1991) (Semenza, G., 2001, Trends Mol. Med., 7, 345-350.Cells are typically cultured in the laboratory at an ambient oxygenconcentration of 21%, but cells in the human body are exposed to muchlower oxygen concentrations ranging from 16% in the lungs to less than6% in most other organs of the body often significantly less in tumors.Semenza G., 2001, Trends Mol. Med., 7, 345-350.

[0003] Solid tumor growth depends on a continuous supply of oxygen andnutrients through neovascularization (angiogenesis). Tumors often becomehypoxic, often because new blood vessels are aberrant and have poorblood flow. Cancer cells make adaptive changes that allow them toproliferate even at hypoxia. These changes include an increase inglycolysis and an increase in production of angiogenic factors. Hypoxiain tumors is associated with resistance to radio- and chemotherapy, andthus is an indicator of poor survival.

[0004] The transcriptional complex, hypoxia inducible factor (HIF), is akey regulator of oxygen homeostasis. Hypoxia induces the expression ofgenes participating in many cellular and physiological processes,including oxygen transport and iron metabolism, erythropoiesis,angiogenesis, glycolysis and glucose uptake, transcription, metabolism,pH regulation, growth-factor signaling, response to stress and celladhesion. These gene products participate in either increasing oxygendelivery to hypoxic tissues or activating an alternative metabolicpathway (glycolysis) which does not require oxygen. Hypoxia-inducedpathways, in addition to being required for normal cellular processes,can also aid tumor growth by allowing or aiding angiogenesis,immortalization, genetic instability, tissue invasion and metastasis(Harris, Nat. Rev. Cancer, 2002, 2, 38-47; Maxwell et al., Curr. Opin.Genet. Dev., 2001, 11, 293-299).

[0005] HIF is a heterodimer composed of an alpha subunit complexed witha beta subunit, both of which are basic helix-loop-helix transcriptionfactors. The beta subunit of HIF is a constitutive nuclear protein. Thealpha subunit is the regulatory subunit specific to the oxygen responsepathway, and can be one of three subunits, HIF1α, 2 alpha or 3 alpha(HIF-1α, HIF-2α and HIF-3α, respectively) (Maxwell et al., Curr. Opin.Genet. Dev., 2001, 11, 293-299; Safran and Kaelin, J. Clin. Invest.,2003, 111, 779-783).

[0006] The transcription factor hypoxia-inducible factor 1 (HIF-1) playsan essential role in homeostatic responses to hypoxia by binding to theDNA sequence 5′-TACGTGCT-3′ and activating the transcription of dozensof genes in vivo under hypoxic conditions (Wang and Semenza, J. Biol.Chem., 1995, 270, 1230-1237). These gene products participate in eitherincreasing oxygen delivery to hypoxic tissues or activating analternative metabolic pathway (glycolysis) which does not requireoxygen. This list includes: aldolase C, enolase 1, glucose transporter1, glucose transporter 3, glyceraldehyde-3-phosphate dehydrogenase,hexokinase 1, hexokinase 2, insulin-like growth factor-2 (IGF-2), IGFbinding protein 1, IGF binding protein 3, lactate dehydrogenase A,phosphoglycerate kinase 1, pyruvate kinase M, p21, transforming growthfactor B3, ceruloplasmin, erythropoietin, transferrin, transferrinreceptor, a1b-adrenergic receptor, adrenomedullin, endothelin-1, hemeoxygenase 1, nitric oxide synthase 2, plasminogen activator inhibitor 1,vascular endothelial growth factor (VEGF), VEGF receptor FTL-1, and p35(Semenza, Genes Dev., 2000, 14, 1983-1991). Expression of HIF1α is alsosensitive to oxygen concentration: increased levels of protein aredetected in cells exposed to 1% oxygen and these decay rapidly uponreturn of the cells to 20% oxygen (Wang et al., Proc. Natl. Acad. Sci.U.S.A., 1995, 92, 5510-5514).

[0007] Hypoxia-inducible factor-1 alpha is a heterodimer composed of a120 kDa alpha subunit complexed with a 91 to 94 kDa beta subunit, bothof which contain a basic helix-loop-helix (Wang and Semenza, J. Biol.Chem., 1995, 270, 1230-1237). The gene encoding hypoxia-induciblefactor-1 alpha (HIF1α, also called HIF-1 alpha, HIF1A, HIF-1A, HIF1-A,and MOP1) was cloned in 1995 (Wang et al., Proc. Natl. Acad. Sci.U.S.A., 1995, 92, 5510-5514). A nucleic acid sequence encoding HIF1α isdisclosed and claimed in U.S. Pat. No. 5,882,914, as are expressionvectors expressing the recombinant DNA, and host cells containing saidvectors (Semenza, 1999).

[0008] HIF1α expression and HIF-1 transcriptional activity are preciselyregulated by cellular oxygen concentration. The beta subunit is aconstitutive nuclear protein, while the alpha subunit is the regulatorysubunit. HIF1α mRNA is expressed at low levels in tissue culture cells,but it is markedly induced by hypoxia or ischemia in vivo (Yu et al., J.Clin. Invest., 1999, 103, 691-696). HIF1α protein is negativelyregulated in non-hypoxic cells by ubiquitination and proteasomaldegradation (Huang et al., Proc. Natl. Acad. Sci. U.S.A., 1998, 95,7987-7992). Under hypoxic conditions, the degradation pathway isinhibited, HIF1α protein levels increase dramatically, and the fractionthat is ubiquitinated decreases. HIF1α then translocates to the nucleusand dimerizes with a beta subunit (Sutter et al., Proc. Natl. Acad. Sci.U.S.A., 2000, 97, 4748-4753).

[0009] A natural antisense transcript that is complementary to the 3′untranslated region of HIF1α mRNA has been discovered and is named“aHIF” (Thrash-Bingham and Tartof, J. Natl. Cancer Inst., 1999, 91,143-151). This is the first case of overexpression of a naturalantisense transcript exclusively associated with a specific humanmalignant disease. aHIF is specifically overexpressed in nonpapillaryclear-cell renal carcinoma under both normoxic and hypoxic conditions,but not in papillary renal carcinoma. Although aHIF is not furtherinduced by hypoxia in nonpapillary disease, it can be induced inlymphocytes where there is a concomitant decrease in HIF1α mRNA.

[0010] HIF1α plays an important role in promoting tumor progression andis overexpressed in common human cancers, including breast, colon, lung,and prostate carcinoma. Overexpression of HIFs is sometimes observed incancers, such as clear cel renal cell carcinoma, even at normoxia.Mutations that inactivate tumor suppressor genes or activate oncogeneshave, as one of their consequences, upregulation of HIF1α activity,either through an increase in HIF1α protein expression, HIF1αtranscriptional activity, or both (Semenza, Pediatr. Res., 2001, 49,614-617).

[0011] Until a tumor establishes a blood supply, the hypoxic conditionslimit tumor growth. Subsequent increases in HIF1α activity result inincreased expression of target genes such as vascular endothelial growthfactor (VEGF). VEGF expression is essential for vascularization and theestablishment of angiogenesis in most solid tumors (Iyer et al., GenesDev., 1998, 12, 149-162). A significant association betweenhypoxia-inducible factor-1 alpha, VEGF overexpression and tumor grade isalso seen in human glioblastoma multiforme, the highest grade glioma inwhich mean patient survival time is less than one year. The rapidlyproliferating tumor outgrows its blood supply, resulting in extensivenecrosis, and these regions express high levels of HIF1α protein andVEGF mRNA, suggesting a response of the tumor to hypoxia (Zagzag et al.,Cancer, 2000, 88, 2606-2618).

[0012] The action of the von Hippel-Landau (VHL) tumor suppressor geneproduct is implicated in hypoxic gene regulation, in both normal anddiseased cells. Individuals with VHL disease are predisposed to renalcysts, clear cell renal carcinoma, phaeochromocytoma, haemangioblastomasof the central nervous system, angiomas of the retina, islet cell tumorsof the pancreas, and endolymphatic sac tumors (Pugh and Ratcliffe,Semin. Cancer. Biol., 2003, 13, 83-89). The VHL gene productparticipates in ubiquitin-mediated proteolysis by acting as therecognition component of the E3-ubiquitin ligase complex involved in thedegradation of hypoxia-inducible factor alpha subunits (Cockman et al.,J. Biol. Chem., 2000, 275, 25733-25741; Ohh et al., Nat. Cell Biol.,2000, 2, 423-427). In normal cells, VHL/HIF complexes form and targetHIF alpha subunits for destruction (Maxwell et al., Nature, 1999, 399,271-275). This is proposed to occur through hydroxylation of theoxygen-dependent domain of HIF2α and subsequent recognition by the VHLgene product, as recognition of a homologous oxygen-dependent domain isthe mechanism by which the VHL protein recognizes HIF1α (Maxwell et al.,Nature, 1999, 399, 271-275). HIF2α is in fact hydroxylated by the enzymeprolyl 4-hydroxylases in vitro (Hirsila et al., J. Biol. Chem., 2003).

[0013] The p53 tumor suppressor also targets HIF1α for degradation bythe proteasome. Loss of p53 activity occurs in the majority of humancancers and indicates that amplification of normal HIF1α levelscontributes to the angiogenic switch during tumorigenesis (Ravi et al.,Genes Dev., 2000, 14, 34-44).

[0014] A mouse model of pulmonary hypertension has shown that localinhibition of HIF1α activity in the lung might represent a therapeuticstrategy for treating or preventing pulmonary hypertension in at riskindividuals. In pulmonary hypertension hypoxia-induced vascularremodeling leads to decreased blood flow, which leads to progressiveright heart failure and death. This hypoxia-induced vascular remodelingis markedly impaired in mice that are partially HIF1α deficient (Yu etal., J. Clin. Invest., 1999, 103, 691-696). Decreased vascular densityand retarded solid tumor growth is also seen in mouse embryonic stemcells which are deficient for HIF1α (Ryan et al., Embo J, 1998, 17,3005-3015).

[0015] During hypoxia, cells shift to a glycolytic metabolic mode fortheir energetic needs and HIF1α is known to upregulate the expression ofmany glycolytic genes. HIF1α may play a pivotal role in the Warburgeffect in tumors, a paradoxical situation in which tumor cells growingunder normoxic conditions show elevated glycolytic rates, which enhancestumor growth and expansion. HIF1α mediates the expression of6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3, a gene whoseprotein product maintains levels of the key regulator of glycolyticflux, fructose-2,6-bisphosphate (Minchenko et al., J. Biol. Chem., 2001,14, 14).

[0016] Currently, there are no known therapeutic agents whicheffectively inhibit the synthesis of HIF1α and to date, investigativestrategies aimed at modulating HIF1α function have involved the use ofantisense expression vectors and oligonucleotides. These studies haveserved to define the involvement of HIF1α in disease progression and toidentify novel roles of HIF1α in vivo including unique roles for HIF1αas a transcription factor under non-hypoxic conditions and as aninhibitor of gene expression.

[0017] Gene transfer of an antisense HIF1α plasmid has been shown toenhance the efficacy of cancer immunotherapy. Antisense therapy wasshown to slow, but not eradicate, the growth of EL-4 tumors establishedin mice. In addition, endogenously expression of HIF1α was almostcompletely inhibited in these tumors. When antisense therapy wascombined with T-cell costimulator B7-1 immunotherapy, the tumorscompletely and rapidly regressed within 1 week. Furthermore, when thesetumor-free mice were rechallenged with EL-4 cells, no tumors emerged,indicating that systemic antitumor immunity had been achieved (Sun etal., Gene Ther., 2001, 8, 638-645).

[0018] Activation of HIF1α is thought to aggravate heart failure byupregulation of cardiac ET-1, a gene product involved in heart failureand whose inhibition improves the survival rate of rats with heartfailure. In a failing heart, a metabolic switch occurs, and HIF1αactivates the expression of glycolytic enzymes as compensation forimpaired b-oxidation of fatty acid. Another consequence of increasedHIF1α activity is that in rat cardiomyocytes, HIF1α was shown to bind tothe 5′-promoter region of the ET-1 gene and increase ET-1 expression. Invitro, an antisense oligonucleotide targeted to hypoxia-induciblefactor-1 alpha largely inhibited the increased gene expression of ET-1,confirming the role of HIF1α in heart failure (Kakinuma et al.,Circulation, 2001, 103, 2387-2394). This antisense oligonucleotide iscomprised of 20 nucleotides and targets bases 11 to 31 of the rat HIF1αwith GenBank accession number AF_(—)057308 incorporated herein byreference.

[0019] Preeclampsia is a disorder of unknown etiology that is theleading cause of fetal and maternal morbidity and mortality. Defectivedownregulation of HIF1α may play a major role in the pathogenesis ofpreeclampsia. For most of the first trimester, the human fetus developsunder hypoxic conditions but at 10-12 weeks the intervillous spaceopens, the fetus is exposed to maternal blood and at this stage thetrophoblast cells invade the maternal decidua. The switch of thetrophoblasts from a proliferative to an invasive phenotype is controlledby cellular oxygen concentration. The proliferative, non-invasivetrophoblast phenotype appears to be maintained by HIF1α mediatedexpression of TGFbeta3 because treatment of human villous explants withan antisense oligonucleotide against HIF1α or TGF beta 3 inducesinvasion under hypoxic conditions. In this case the HIF1α antisenseoligonucelotide was comprised of phosphorothioate oligonucleotides, 16nucleotides in length, and targeted to the AUG codon (Caniggia et al.,J. Clin. Invest., 2000, 105, 577-587.; Caniggia et al., Placenta, 2000,21 Suppl A, S25-30).

[0020] The human intestinal trefoil factor (ITF) gene product protectsthe epithelial barrier during episodes of intestinal hypoxia. The ITFgene promoter bears a bindingsite for hypoxia-inducible factor-1 alpha,and the function of HIF1α as a transcription factor for ITF wasconfirmed in vitro. T84 colonic epithelial cells were treated with aphosphorothioate antisense oligonucleotide, 15 nucleotides in length andtargeted to the AUG codon of HIF1α and this resulted in a loss of ITFhypoxia inducibility (Furuta et al., J. Exp. Med., 2001, 193,1027-1034).

[0021] Human epidemiological and animal studies have associatedinhalation of nickel dusts with an increased incidence of pulmonaryfibrosis. Nickel transcriptionally activates plasminogen activatorinhibitor (PAI-1), an inhibitor of fibrinolysis, through the HIF1αsignaling pathway. This was evidenced by decreases in PAI-1 mRNA levelswhen human airway epithelial cells were treated with an antisenseoligonucleotide directed against HIF1α identical to the one used in thepreeclampsia study discussed above. These data may be critical forunderstanding the pathology of pulmonary fibrosis and other diseasesassociated with nickel exposure (Andrew et al., Am J Physiol Lung CellMol Physiol, 2001, 281, L607-615).

[0022] HIF1α is constitutively expressed in cerebral neurons undernormoxic conditions. A second dimerization partner for HIF1α is ARNT2, acerebral translocator homologous to hypoxia-inducible factor-1 beta. Onesplice variant of HIF1α found in rat neurons dimerizes with ARNT2 moreavidly than it does with HIF1b, and the resulting hypoxia-induciblefactor-1 alpha-ARNT2 heterodimer does not recognize the HIF1α bindingsite of the erythropoietin gene. This suggests that transcription of adifferent set of genes is controlled by the hypoxia-inducible factor-1alpha-ARNT2 heterodimer controls in neurons under nonhypoxic conditionsthan the hypoxia-inducible factor-1 alpha-HIF1α heterodimer controlsunder hypoxic conditions. This was evidenced by antisenseoligonucleotide downregulation of HIF1α expression in which theantisense oligonucleotide consisted of 16 phosphorothioate nucleotidestargeted to bases 38 to 54 of the rat hypoxia-inducible factor-1 withGenBank accession number AF_(—)057308 (Drutel et al., Eur. J. Neurosci.,2000, 12, 3701-3708).

[0023] A role for HIF1α in mediating a down-regulatory pathway wasrecently discovered using antisense oligonucleotide depletion ofhypoxia-inducible factor-1 alpha. The peroxisome proliferator-activatedreceptors (PPARS) are a nuclear hormone-binding proteins that regulatetranscriptional activities. Ligands which bind the PPAR-gamma isoformman amplify or inhibit the expression of inflammation-related geneproducts and may regulate the duration of inflammatory response. Hypoxiaelicits a down-regulation of PPAR-gamma in intestinal epithelial cellswhich is effected through a binding site for HIF1α on the antisensestrand of the PPAR-gamma gene. The expression of PPAR-gamma wasupregulated in hypoxic cells when treated with an antisenseoligonucleotide targeted to HIF1α identical to the one used in thepreeclampsia study discussed above (Narravula and Colgan, J. Immunol.,2001, 166, 7543-7548).

[0024] The gene encoding hypoxia-inducible factor 2 alpha (HIF2α; alsocalled HIF-2 alpha, endothelial PAS domain protein 1, EPAS1, MOP2,hypoxia-inducible factor 2, HIF-related factor, HRF, HIF1 alpha-likefactor, HLF) was initially identified as a transcription factorexpressed in endothelial cells (Ema et al., Proc. Natl. Acad. Sci.U.S.A., 1997, 94, 4273-4278; Flamme et al., Mech. Dev., 1997, 63, 51-60;Hogenesch et al., J. Biol. Chem., 1997, 272, 8581-8593; Tian et al.,Genes Dev., 1997, 11, 72-82). A nucleic acid sequence encoding humanHIF2α is disclosed and claimed in U.S. Pat. No. 5,695,963 (McKnight etal., 1997).

[0025] HIF2α mRNA is primarily expressed in highly vascularized adulttissues, such as lung, heart and liver, and in the placenta andendothelial cells of the embryonic and adult mouse (Hogenesch et al., J.Biol. Chem., 1997, 272, 8581-8593). Comparison of normal human tissuesand cancers reveals that HIF2α protein is not detectable in normaltissue, but is easily visualized in malignant tissues (Talks et al., Am.J. Pathol., 2000, 157, 411-421). The requirement for expression of HIF2αin development is demonstrated by the abnormalities observed in HIF2αgene deficient mouse embyros, which include the disruption ofcatecholamine homeostasis and lack of protection against heart failureobserved (Tian et al., Genes Dev., 1998, 12, 3320-3324). Targeteddisruption of the HIF2α gene and generation of embryos deficient forHIF2α is disclosed in the PCT publication WO 02/086497 (Compernolle etal., 2002). This publication also discloses antisenseoligodeoxyribonucleotides for use in inhibiting HIF2α expressiontargeted to the translation initiation codon of HIF2α (Compernolle etal., 2002).

[0026] HIF2α expression and HIF transcriptional activity are preciselyregulated by cellular oxygen concentration. Whereas changes in oxygenlevels do not affect HIF1-beta protein levels, the abundance of thealpha subunits is markedly increased upon exposure of cells to hypoxia,primarily due to stabilization of the alpha subunit protein (Safran andKaelin, J. Clin. Invest., 2003, 111, 779-783). HIF2α mRNA and protein isexpressed at low levels in tissue culture cells, but protein expressionis markedly induced by exposure to 1% oxygen, a hypoxic state (Wieseneret al., Blood, 1998, 92, 2260-2268). The hypoxia-inducible factor 2alpha/hypoxia-inducible factor 1 beta heterodimer protein binds to thehypoxic response element, which contains the core recognition sequence5′-TACGTG-3′ and is found in the cis-regulatory regions ofhypoxia-regulated genes (Ema et al., Proc. Natl. Acad. Sci. U.S.A.,1997, 94, 4273-4278; Hogenesch et al., J. Biol. Chem., 1997, 272,8581-8593). Binding of the heterodimer to the HRE induces geneexpression. Upon return to normoxic conditions, HIF2α protein is rapidlydegraded (Wiesener et al., Blood, 1998, 92, 2260-2268).

[0027] The mitogen-activated protein kinase (MAPK) pathway is criticalfor HIF2α activation. Inhibition of a dual specificity protein kinasethat directly phosphorylates MAPK prevents HIF2α trans-activation duringhypoxia (Conrad 1999; Conrad, 2001). However, the inhibitor does notprevent HIF2α phosphorylation, thus, while the MAPK pathway regulatesthe activity of hypoxia-inducible factor 2 alpha, it does not directlyphosphorylate the protein (Conrad et al., Comp. Biochem. Physiol. B.Biochem. Mol Biol., 2001, 128, 187-204; Conrad et al., J. Biol. Chem.,1999, 274, 33709-33713). The Src family kinase pathway is alsoimplicated in regulation of hypoxia-inducible factor 2 alpha. A specificinhibitor of the Src family of kinases abolishes the hypoxia-inducedexpression of HIF2α mRNA in human lung adenocarcinoma cells (Sato etal., Am. J. Respir. Cell Mol. Biol., 2002, 26, 127-134).

[0028] The maintenance of oxygen homeostasis, in addition to beingrequired in physiological development, is also required in tumor growth.Tumor cells experience hypoxia because blood circulates poorly throughthe aberrant blood vessel that tumors establish. Although hypoxia istoxic to cancer cells, they survive as a result of genetic and adaptivechanges that allow them to thrive in a hypoxic environment. One suchadaptation is an increase in the expression of the angiogenic growthfactor named vascular endothelial growth factor (VEGF). VEGF is a keyangiogenic factor secreted by cancer cells, as well as normal cells, inresponse to hypoxia (Harris, Nat. Rev. Cancer, 2002, 2, 38-47; Maxwellet al., Curr. Opin. Genet. Dev., 2001, 11, 293-299).

[0029] Hemangioblastomas, the most frequent manifestation of VHL genemutations, exhibit overexpression of VEGF mRNA in their associatedstromal cells. The VEGF mRNA overexpression is highly correlated withelevated expression of HIF2α mRNA. This finding suggests a relationshipbetween loss of function of the VHL gene, and transcriptional activationof the VEGF gene, possibly through HIF2α activity in VEGF-dependentvascular growth (Flamme et al., Am. J. Pathol., 1998, 153, 25-29).

[0030] The tumor suppressive activity of the VHL gene product can beoverridden by the activation of HIF target genes in human renalcarcinoma cells in vivo. VHL gene product mutants lose the ability totarget HIF for ubiquitin-mediated destruction, suggesting that downregulation of HIF and VHL tumor suppressor function are intimatelylinked (Kondo et al., Cancer Cell, 2002, 1, 237-246). In contrast tohuman renal cell carcinoma, the product of the tuberous sclerosiscomplex-2 (Tsc-2) gene, product rather than VHL gene, is the primarytarget for rodent renal cell carcinoma (Liu et al., Cancer Res., 2003,63, 2675-2680). Rat RCC cells lacking Tsc-2 function exhibitstabilization of HIF2α protein and upregulation of VEGF, and were highlyvascularized (Liu et al., Cancer Res., 2003, 63, 2675-2680).

[0031] A link between elevated HIF2α activity and angiogenesis has alsobeen demonstrated by experiments that show how HIF activity regulatesVEGF expression. Normal human kidney cells typically have low levels ofhypoxia-inducible factor 2 alpha, but upon introduction of a vectorencoding HIF2α into these cells, VEGF mRNA and protein levels increasesignificantly (Xia et al., Cancer, 2001, 91, 1429-1436). When HIF2α wasinhibited, VEGF expression was significantly decreased, thusdemonstrating a direct link between HIF2αactivity and VEGF expression(Xia et al., Cancer, 2001, 91, 1429-1436). Similarly, a dose-dependentincrease in VEGF mRNA is observed when human umbulical vein cells aretransduced with a virus encoding HIF2α (Maemura et al., J. Biol. Chem.,1999, 274, 31565-31570). Expression of a mutated HIF2α that lacks atransactivation domain inhibits the induction of VEGF mRNA duringhypoxia, a finding that further suggests that HIF2α is an importantregulator of VEGF expression (Maemura et al., J. Biol. Chem., 1999, 274,31565-31570).

[0032] A correlation between HIF activity and VEGF expression is alsoobserved in malignant cells and tissues. HIF2α can be readily detectedin renal cell carcinoma (RCC) cell lines in the absence of a vectorencoding HIF2α (Xia et al., Cancer, 2001, 91, 1429-1436). Significantincreases in HIF2α and VEGF mRNA in renal cell carcinoma tissue samples,compared to normal tissue, suggest that abnormal activation of HIF2α maybe involved in the angiogenesis of RCC (Xia et al., Cancer, 2001, 91,1429-1436).

[0033] In addition to RCC, the expression of HIF2α in other malignancieshas also been reported. HIF2α is expressed at the levels of mRNA andprotein in human bladder cancers, especially in those with an invasivephenotype (Xia et al., Urology, 2002, 59, 774-778). Another example ofoverexpression of HIF2α is seen in squamous cell head-and-neck cancer(SCHNC). Higher levels of HIF2α were associated with locally aggressivebehavior of SCHNC, as well as intensification of angiogenesis(Koukourakis et al., Int. J. Radiat. Oncol. Biol. Phys., 2002, 53,1192-1202). These findings also demonstrated a link betweenoverexpression of HIF2α and resistance to chemotherapy. Yet anothercorrelation between overexpression of HIF2α and cancer is seen inmalignant pheochromocytomas, which exhibit a higher level of HIF2α andan induced VEGF pathway, when compared to benign counterparts (Favier etal., Am. J. Pathol., 2002, 161, 1235-1246). HIF2α overexpression is alsoa common event in non-small-cell lung cancer (NSCLC) and is related tothe up-regulation of multiple angiogenic factors and overexpression ofangiogenic receptors by cancer cells. HIF2α overexpression in NSCLC isan indicator of poor prognosis (Giatromanolaki et al., Br. J. Cancer,2001, 85, 881-890). Taken together, these studies demonstrate thatelevated HIF2α confers aggressive tumor behavior, and that targeting theHIF pathway may aid the treatment of several different types of cancers.

[0034] Overexpression of HIF2α has also been observed in several cancercell lines in addition to RCC cell lines. Elevated levels of HIF2α mRNAand protein are seen in human lung adenocarcinoma cells, and exposure ofthese cells to hypoxia further increases HIF2α expression (Sato et al.,Am. J. Respir. Cell Mol. Biol., 2002, 26, 127-134). Furthermore, thehypoxia response element plays a role in constitutively upregulating anisoform of VEGF in cancer cell lines under normoxic conditions. The HRElocated within a cell type-specific enhancer element in glioblastomacells participates in the up-regulation of VEGF expression throughenhanced binding of HIF2α to the HRE (Liang et al., J. Biol. Chem.,2002, 277, 20087-20094). A truncated version of HIF2α that can bind tohypoxia-inducible factor 1 beta, but not to the HRE, was unable totransactivate the VEGF promoter (Liang et al., J. Biol. Chem., 2002,277, 20087-20094). This further demonstrates the capability of cancercells to combat hypoxic conditions by enhancing expression of factorsrequired for vascularization and angiogenesis.

[0035] Short interfering RNAs (siRNAs) have been used to specificallyinhibit the expression of HIF1α and HIF2α in human breast and renalcarcinoma cell lines and in a human endothelial cell line. SiRNAduplexes with dTdT overhangs at both ends were designed to targetnucleotides 1521-1541 and 1510-1530 of the HIF1α mRNA squence (NM001530)and nucleotides 1260-1280 and 328-348 of the HIF2α sequence (NM001430).It was found that in the breast carcinoma and endothelial cell lines,gene expression and cell migration patterns were critically dependent onHIF1α but not hypoxia-inducible factor-2 alpha, but critically dependenton HIF2α in the renal carcinoma cells. Sowter et al., 2003, Cancer Res.,63, 6130-6134.

[0036] Defective downregulation of HIF2α may play a major role in thepathogenesis of preeclampsia. HIF2α protein levels are increased duringearly development, as expected in a hypoxic environment, and thendecrease significantly with gestational age (Rajakumar and Conrad, Biol.Reprod., 2000, 63, 559-569). However, HIF2α protein expression issignificantly increased in preeclamptic relative to normal termplacentas (Rajakumar et al., Biol. Reprod., 2001, 64, 499-506). Thisresult suggests that failure to down-regulate HIF2α protein expressionduring early pregnancy could prevent the switch of the trophoblast to aninvasive phenotype and ultimately lead to preeclampsia (Rajakumar etal., Biol. Reprod., 2001, 64, 499-506).

[0037] Overexpression of hypoxia-inducible factor 2 alpha, as well ashypoxia-inducible factor 1, has been observed in the inflammatory boweldiseases Crohn's disease and ulcerative colitis (Giatromanolaki et al.,J. Clin. Pathol., 2003, 56, 209-213). However, VEGF expression was weakin ulcerative colitis samples, and absent in Crohn's disease samples.The discordant expression of VEGF and HIF2α may lead to a reducedability of a tissue to produce or respond to VEGF, which may in turnlead to reduced endothelial and epithelial cell viability(Giatromanolaki et al., J. Clin. Pathol., 2003, 56, 209-213).

[0038] In addition to participating in adaptive changes in response tohypoxia, HIF2α may also function in an inflammatory response in cardiacmyocytes. In cultured cardiac myocytes, interleukin-1 beta (IL-1beta)significantly increased both HIF2α mRNA and protein levels (Tanaka etal., J. Mol. Cell Cardiol., 2002, 34, 739-748). Transduction of cardiacmyocytes with adenovirus expressing HIF2α dramatically increased thelevels of adrenomedullin (AM) mRNA, which is also upregulated byIL-lbeta (Tanaka et al., J. Mol. Cell Cardiol., 2002, 34, 739-748).Since IL-1 beta has been implicated in the pathogenesis of heartfailure, and AM is known to improve cardiac function during heartfailure, these results suggest that HIF2α plays a role in the adaptationof the cardiac myocytes during heart failure (Tanaka et al., J. Mol.Cell Cardiol., 2002, 34, 739-748).

[0039] Disclosed and claimed in the PCT publication WO 00/09657 is amethod of inhibiting angiogenesis in a mammal through administration ofa compound which inhibits the binding of human HIF2α protein to the DNAregulatory element of an angiogenic factor, wherein the compound can bean antisense nucleic acid molecule complementary to all or part of themRNA encoding human HIF2α (Lee et al., 2000). This publication alsodiscloses a nucleic acid encoding human hypoxia-inducible factor 2alpha.

[0040] The PCT publication WO 01/62965 discloses and claims adifferential screening method for identifying a genetic element involvedin a cellular process, which method includes introducing HIF2α intocells (Kingsman, 2001). This publication also discloses the developmentof HIF2α agonists or antagonists.

[0041] The PCT publication WO 02/34291 claims methods and reagents,including the use of antisense oligonucleotides, for the inhibition ofhuman HIF1α transcription (Colgan, 2002). This publication alsodiscloses a nucleic acid encoding human hypoxia-inducible factor 2alpha.

[0042] U.S. Pat. No. 6,395,548 claims a nucleic acid encoding a deletionmutant of human HIF2α and the use of this deletion mutant as a method ofinhibiting expression of an angiogenic factor in vitro. This patent alsodiscloses a nucleic acid encoding human hypoxia-inducible factor 2alpha, as well as nucleic acids complementary to all or part of thehuman HIF2α cDNA for use in antisense treatment to inhibit theexpression of HIF2α (Lee et al., 2002).

[0043] U.S. Pat. No. 6,432,927 discloses nucleic acid sequences,including sense and antisense oligonucleotides, which are derived froman HIF2α and incorporated into recombinant nucleic acid molecules forthe purpose of sustaining HIF2α expression in cells (Gregory andVincent, 2002).

[0044] The nucleic acid sequence encoding a human HIF2α and insertion ofthis sequence into a viral expression vector, for the purpose of drivinghuman HIF2α expression in mammalian cells, is disclosed in the PCTpublication WO 02/068466 (White et al., 2002).

[0045] The PCT publication WO 02/094862 discloses a method forintroducing into a muscle cell a nucleic acid sequence encodinghypoxia-inducible factor 2 alpha, for the purpose of overexpressingHIF2α and stimulating angiogenesis or metabolic activity (Guy, 2002).

[0046] Disclosed and claimed in the US pre-grant publication2003/0045686 is a nucleic acid encoding human hypoxia-inducible factor 2alpha, and a method of delivering a therapeutically effective amount ofthis nucleic acid to a subject for the purpose of reducing or preventinghypoxia (Kaelin Jr. and Ivan, 2003). This publication also discloses andclaims human HIF muteins, including HIF2α mutein, which are designed tobe more stable and/or resistant to degradation.

[0047] As a consequence of HIF2α involvement in many diseases, thereremains a long felt need for additional agents capable of effectivelyregulating HIF2α function. As such, inhibition is especially importantin the treatment of cancer, given that the upregulation of expression ofHIF2α is associated with so many different types of cancer.

[0048] As a consequence of HIF1α and HIF2α involvement in many diseases,there remains a long felt need for additional agents capable ofeffectively inhibiting HIF1α and HIF2α function.

[0049] Antisense technology is emerging as an effective means forreducing the expression of specific gene products and may thereforeprove to be uniquely useful in a number of therapeutic, diagnostic, andresearch applications for the modulation of HIF1α and HIF2α expression.

[0050] The present invention provides compositions and methods formodulating HIF1α and HIF2α expression. In particular antisensecompositions for modulating HIF1α and/or HIF2α expression are believedto be useful in treatment of abnormal proliferative conditionsassociated with HIF1α and/or HIF2α. Examples of abnormal proliferativeconditions are hyperproliferative disorders such as cancers, tumors,hyperplasias, pulmonary fibrosis, angiogenesis, psoriasis,atherosclerosis and smooth muscle cell proliferation in the bloodvessels, such as stenosis or restenosis following antioplasty. It ispresently believed that inhibition of both HIF1α and HIF2α may be aparticularly useful approach to treatment of such disorders.

SUMMARY OF THE INVENTION

[0051] The present invention is directed to compounds, especiallynucleic acid and nucleic acid-like oligomers, which are targeted to anucleic acid encoding HIF1α and/or HIF2α, and which modulate theexpression of HIF1α and/or HIF2α. Pharmaceutical and other compositionscomprising the compounds of the invention are also provided. Furtherprovided are methods of screening for modulators of HIF1α and/or HIF2αand methods of modulating the expression of HIF1α and/or HIF2α in cells,tissues or animals comprising contacting said cells, tissues or animalswith one or more of the compounds or compositions of the invention.Methods of treating an animal, particularly a human, suspected of havingor being prone to a disease or condition associated with expression ofHIF1α and/or HIF2α are also set forth herein. Such methods compriseadministering a therapeutically or prophylactically effective amount ofone or more of the compounds or compositions of the invention to theperson in need of treatment.

DETAILED DESCRIPTION OF THE INVENTION

[0052] A. Overview of the Invention

[0053] The present invention employs compounds, preferablyoligonucleotides and similar species for use in modulating the functionor effect of nucleic acid molecules encoding HIF1α or HIF2α. This isaccomplished by providing oligonucleotides which specifically hybridizewith one or more nucleic acid molecules encoding HIF1α or HIF2α. Thus“target nucleic acid” refers to a nucleic acid molecule encoding HIF1αor HIF2α. As used herein, the term “nucleic acid molecule encodingHIF1α” has been used for convenience to encompass DNA encoding HIF1α,RNA (including pre-mRNA and mRNA or portions thereof) transcribed fromsuch DNA, and also cDNA derived from such RNA. Similarly, the term“nucleic acid molecule encoding HIF2α” has been used for convenience toencompass DNA encoding HIF2α, RNA (including pre-mRNA and mRNA orportions thereof) transcribed from such DNA, and also cDNA derived fromsuch RNA. The hybridization of a compound of this invention with itstarget nucleic acid is generally referred to as “antisense”.Consequently, the preferred mechanism believed to be included in thepractice of some preferred embodiments of the invention is referred toherein as “antisense inhibition.” Such antisense inhibition is typicallybased upon hydrogen bonding-based hybridization of oligonucleotidestrands or segments such that at least one strand or segment is cleaved,degraded, or otherwise rendered inoperable. In this regard, it ispresently preferred to target specific nucleic acid molecules and theirfunctions for such antisense inhibition.

[0054] The functions of DNA to be interfered with can includereplication and transcription. Replication and transcription, forexample, can be from an endogenous cellular template, a vector, aplasmid construct or otherwise. The functions of RNA to be interferedwith can include functions such as translocation of the RNA to a site ofprotein translation, translocation of the RNA to sites within the cellwhich are distant from the site of RNA synthesis, translation of proteinfrom the RNA, splicing of the RNA to yield one or more RNA species, andcatalytic activity or complex formation involving the RNA which may beengaged in or facilitated by the RNA. One preferred result of suchinterference with target nucleic acid function is modulation of theexpression of HIF1α or HIF2α. In the context of the present invention,“modulation” and “modulation of expression” mean either an increase(stimulation) or a decrease (inhibition) in the amount or levels of anucleic acid molecule encoding the gene, e.g., DNA or RNA. Inhibition isoften the preferred form of modulation of expression and mRNA is often apreferred target nucleic acid.

[0055] In the context of this invention, “hybridization” means thepairing of complementary strands of oligomeric compounds. In the presentinvention, the preferred mechanism of pairing involves hydrogen bonding,which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogenbonding, between complementary nucleoside or nucleotide bases(nucleobases) of the strands of oligomeric compounds. For example,adenine and thymine are complementary nucleobases which pair through theformation of hydrogen bonds. Hybridization can occur under varyingcircumstances.

[0056] An antisense compound is specifically hybridizable when bindingof the compound to the target nucleic acid interferes with the normalfunction of the target nucleic acid to cause a loss of activity, andthere is a sufficient degree of complementarity to avoid non-specificbinding of the antisense compound to non-target nucleic acid sequencesunder conditions in which specific binding is desired, i.e., underphysiological conditions in the case of in vivo assays or therapeutictreatment, and under conditions in which assays are performed in thecase of in vitro assays.

[0057] In the present invention the phrase “stringent hybridizationconditions” or “stringent conditions” refers to conditions under which acompound of the invention will hybridize to its target sequence, but toa minimal number of other sequences. Stringent conditions aresequence-dependent and will be different in different circumstances andin the context of this invention, “stringent conditions” under whicholigomeric compounds hybridize to a target sequence are determined bythe nature and composition of the oligomeric compounds and the assays inwhich they are being investigated.

[0058] “Complementary,” as used herein, refers to the capacity forprecise pairing between two nucleobases of an oligomeric compound. Forexample, if a nucleobase at a certain position of an oligonucleotide (anoligomeric compound), is capable of hydrogen bonding with a nucleobaseat a certain position of a target nucleic acid, said target nucleic acidbeing a DNA, RNA, or oligonucleotide molecule, then the position ofhydrogen bonding between the oligonucleotide and the target nucleic acidis considered to be a complementary position. The oligonucleotide andthe further DNA, RNA, or oligonucleotide molecule are complementary toeach other when a sufficient number of complementary positions in eachmolecule are occupied by nucleobases which can hydrogen bond with eachother. Thus, “specifically hybridizable” and “complementary” are termswhich are used to indicate a sufficient degree of precise pairing orcomplementarity over a sufficient number of nucleobases such that stableand specific binding occurs between the oligonucleotide and a targetnucleic acid.

[0059] It is understood in the art that the sequence of an antisensecompound need not be 100% complementary to that of its target nucleicacid to be specifically hybridizable. Moreover, an oligonucleotide mayhybridize over one or more segments such that intervening or adjacentsegments are not involved in the hybridization event (e.g., a loopstructure or hairpin structure). It is preferred that the antisensecompounds of the present invention comprise at least 70% sequencecomplementarity to a target region within the target nucleic acid, morepreferably that they comprise 90% sequence complementarity and even morepreferably comprise 95% sequence complementarity to the target regionwithin the target nucleic acid sequence to which they are targeted. Forexample, an antisense compound in which 18 of 20 nucleobases of theantisense compound are complementary to a target region, and wouldtherefore specifically hybridize, would represent 90 percentcomplementarity. In this example, the remaining noncomplementarynucleobases may be clustered or interspersed with complementarynucleobases and need not be contiguous to each other or to complementarynucleobases. As such, an antisense compound which is 18 nucleobases inlength having 4 (four) noncomplementary nucleobases which are flanked bytwo regions of complete complementarity with the target nucleic acidwould have 77.8% overall complementarity with the target nucleic acidand would thus fall within the scope of the present invention. Percentcomplementarity of an antisense compound with a region of a targetnucleic acid can be determined routinely using BLAST programs (basiclocal alignment search tools) and PowerBLAST programs known in the art(Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden,Genome Res., 1997, 7, 649-656).

[0060] B. Compounds of the Invention

[0061] According to the present invention, compounds include antisenseoligomeric compounds, antisense oligonucleotides, ribozymes, externalguide sequence (EGS) oligonucleotides, alternate splicers, primers,probes, and other oligomeric compounds which hybridize to at least aportion of the target nucleic acid. As such, these compounds may beintroduced in the form of single-stranded, double-stranded, circular orhairpin oligomeric compounds and may contain structural elements such asinternal or terminal bulges or loops. Once introduced to a system, thecompounds of the invention may elicit the action of one or more enzymesor structural proteins to effect modification of the target nucleicacid.

[0062] One non-limiting example of such an enzyme is RNAse H, a cellularendonuclease which cleaves the RNA strand of an RNA:DNA duplex. It isknown in the art that single-stranded antisense compounds which are“DNA-like” elicit RNAse H. Activation of RNase H, therefore, results incleavage of the RNA target, thereby greatly enhancing the efficiency ofoligonucleotide-mediated inhibition of gene expression. Similar roleshave been postulated for other ribonucleases such as those in the RNaseIII and ribonuclease L family of enzymes.

[0063] While the preferred form of antisense compound is asingle-stranded antisense oligonucleotide, in many species theintroduction of double-stranded structures, such as double-stranded RNA(dsRNA) molecules, has been shown to induce potent and specificantisense-mediated reduction of the function of a gene or its associatedgene products. This phenomenon occurs in both plants and animals and isbelieved to have an evolutionary connection to viral defense andtransposon silencing.

[0064] The first evidence that dsRNA could lead to gene silencing inanimals came in 1995 from work in the nematode, Caenorhabditis elegans(Guo and Kempheus, Cell, 1995; 81, 611-620). Montgomery et al. haveshown that the primary interference effects of dsRNA areposttranscriptional (Montgomery et al., Proc. Natl. Acad. Sci. USA,1998, 95, 15502-15507). The posttranscriptional antisense mechanismdefined in Caenorhabditis elegans resulting from exposure todouble-stranded RNA (dsRNA) has since been designated RNA interference(RNAi). This term has been generalized to mean antisense-mediated genesilencing involving the introduction of dsRNA leading to thesequence-specific reduction of endogenous targeted mRNA levels (Fire etal., Nature, 1998, 391, 806-811). Recently, it has been shown that itis, in fact, the single-stranded RNA oligomers of antisense polarity ofthe dsRNAs which are the potent inducers of RNAi (Tijsterman et al.,Science, 2002, 295, 694-697).

[0065] The oligonucleotides of the present invention also includevariants in which a different base is present at one or more of thenucleotide positions in the oligonucleotide. For example, if the firstnucleotide is an adenosine, variants may be produced which containthymidine, guanosine or cytidine at this position. This may be done atany of the positions of the oligonucleotide. These oligonucleotides arethen tested using the methods described herein to determine theirability to inhibit expression of HIF2α mRNA.

[0066] In the context of this invention, the term “oligomeric compound”refers to a polymer or oligomer comprising a plurality of monomericunits. In the context of this invention, the term “oligonucleotide”refers to an oligomer or polymer of ribonucleic acid (RNA) ordeoxyribonucleic acid (DNA) or mimetics, chimeras, analogs and homologsthereof. This term includes oligonucleotides composed of naturallyoccurring nucleobases, sugars and covalent internucleoside (backbone)linkages as well as oligonucleotides having non-naturally occurringportions which function similarly. Such modified or substitutedoligonucleotides are often preferred over native forms because ofdesirable properties such as, for example, enhanced cellular uptake,enhanced affinity for a target nucleic acid and increased stability inthe presence of nucleases.

[0067] While oligonucleotides are a preferred form of the compounds ofthis invention, the present invention comprehends other families ofcompounds as well, including but not limited to oligonucleotide analogsand mimetics such as those described herein.

[0068] The compounds in accordance with this invention preferablycomprise from about 8 to about 80 nucleobases (i.e. from about 8 toabout 80 linked nucleosides). One of ordinary skill in the art willappreciate that the invention embodies compounds of 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66,67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 nucleobases inlength.

[0069] In one preferred embodiment, the compounds of the invention are12 to 50 nucleobases in length. One having ordinary skill in the artwill appreciate that this embodies compounds of 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50nucleobases in length.

[0070] In another preferred embodiment, the compounds of the inventionare 15 to 30 nucleobases in length. One having ordinary skill in the artwill appreciate that this embodies compounds of 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length.

[0071] Particularly preferred compounds are oligonucleotides from about12 to about 50 nucleobases, even more preferably those comprising fromabout 15 to about 30 nucleobases.

[0072] Antisense compounds 8-80 nucleobases in length comprising astretch of at least eight (8) consecutive nucleobases selected fromwithin the illustrative antisense compounds are considered to besuitable antisense compounds as well.

[0073] Exemplary preferred antisense compounds include oligonucleotidesequences that comprise at least the 8 consecutive nucleobases from the5′-terminus of one of the illustrative preferred antisense compounds(the remaining nucleobases being a consecutive stretch of the sameoligonucleotide beginning immediately upstream of the 5′-terminus of theantisense compound which is specifically hybridizable to the targetnucleic acid and continuing until the oligonucleotide contains about 8to about 80 nucleobases). Similarly preferred antisense compounds arerepresented by oligonucleotide sequences that comprise at least the 8consecutive nucleobases from the 3′-terminus of one of the illustrativepreferred antisense compounds (the remaining nucleobases being aconsecutive stretch of the same oligonucleotide beginning immediatelydownstream of the 3′-terminus of the antisense compound which isspecifically hybridizable to the target nucleic acid and continuinguntil the oligonucleotide contains about 8 to about 80 nucleobases). Onehaving skill in the art armed with the preferred antisense compoundsillustrated herein will be able, without undue experimentation, toidentify further preferred antisense compounds.

[0074] C. Targets of the Invention

[0075] “Targeting” an antisense compound to a particular nucleic acidmolecule, in the context of this invention, can be a multistep process.The process usually begins with the identification of a target nucleicacid whose function is to be modulated. This target nucleic acid may be,for example, a cellular gene (or mRNA transcribed from the gene) whoseexpression is associated with a particular disorder or disease state, ora nucleic acid molecule from an infectious agent. In the presentinvention, the target nucleic acid encodes HIF1α or HIF2α.

[0076] The targeting process usually also includes determination of atleast one target region, segment, or site within the target nucleic acidfor the antisense interaction to occur such that the desired effect,e.g., modulation of expression, will result. Within the context of thepresent invention, the term “region” is defined as a portion of thetarget nucleic acid having at least one identifiable structure,function, or characteristic. Within regions of target nucleic acids aresegments. “Segments” are defined as smaller or sub-portions of regionswithin a target nucleic acid. “Sites,” as used in the present invention,are defined as positions within a target nucleic acid.

[0077] Since, as is known in the art, the translation initiation codonis typically 5′-AUG (in transcribed mRNA molecules; 5′-ATG in thecorresponding DNA molecule), the translation initiation codon is alsoreferred to as the “AUG codon,” the “start codon” or the “AUG startcodon”. A minority of genes have a translation initiation codon havingthe RNA sequence 5′-GUG, 5′-UUG or 5′-CUG, and 5′-AUA, 5′-ACG and 5′-CUGhave been shown to function in vivo. Thus, the terms “translationinitiation codon” and “start codon” can encompass many codon sequences,even though the initiator amino acid in each instance is typicallymethionine (in eukaryotes) or formylmethionine (in prokaryotes). It isalso known in the art that eukaryotic and prokaryotic genes may have twoor more alternative start codons, any one of which may be preferentiallyutilized for translation initiation in a particular cell type or tissue,or under a particular set of conditions. In the context of theinvention, “start codon” and “translation initiation codon” refer to thecodon or codons that are used in vivo to initiate translation of an mRNAtranscribed from a gene encoding HIF1α or HIF2α, regardless of thesequence(s) of such codons. It is also known in the art that atranslation termination codon (or “stop codon”) of a gene may have oneof three sequences, i.e., 5′-UAA, 5′-UAG and 5′-UGA (the correspondingDNA sequences are 5′-TAA, 5′-TAG and 5′-TGA, respectively).

[0078] The terms “start codon region” and “translation initiation codonregion” refer to a portion of such an mRNA or gene that encompasses fromabout 25 to about 50 contiguous nucleotides in either direction (i.e.,5′ or 3′) from a translation initiation codon. Similarly, the terms“stop codon region” and “translation termination codon region” refer toa portion of such an mRNA or gene that encompasses from about 25 toabout 50 contiguous nucleotides in either direction (i.e., 5′ or 3′)from a translation termination codon. Consequently, the “start codonregion” (or “translation initiation codon region”) and the “stop codonregion” (or “translation termination codon region”) are all regionswhich may be targeted effectively with the antisense compounds of thepresent invention.

[0079] The open reading frame (ORF) or “coding region,” which is knownin the art to refer to the region between the translation initiationcodon and the translation termination codon, is also a region which maybe targeted effectively. Within the context of the present invention, apreferred region is the intragenic region encompassing the translationinitiation or termination codon of the open reading frame (ORF) of agene.

[0080] Other target regions include the 5′ untranslated region (5′UTR),known in the art to refer to the portion of an mRNA in the 5′ directionfrom the translation initiation codon, and thus including nucleotidesbetween the 5′ cap site and the translation initiation codon of an mRNA(or corresponding nucleotides on the gene), and the 3′ untranslatedregion (3′UTR), known in the art to refer to the portion of an mRNA inthe 3′ direction from the translation termination codon, and thusincluding nucleotides between the translation termination codon and 3′end of an mRNA (or corresponding nucleotides on the gene). The 5′ capsite of an mRNA comprises an N7-methylated guanosine residue joined tothe 5′-most residue of the mRNA via a 5′-5′ triphosphate linkage. The 5′cap region of an mRNA is considered to include the 5′ cap structureitself as well as the first 50 nucleotides adjacent to the cap site. Itis also preferred to target the 5′ cap region.

[0081] Although some eukaryotic mRNA transcripts are directlytranslated, many contain one or more regions, known as “introns,” whichare excised from a transcript before it is translated. The remaining(and therefore translated) regions are known as “exons” and are splicedtogether to form a continuous mRNA sequence. Targeting splice sites,i.e., intron-exon junctions or exon-intron junctions, may also beparticularly useful in situations where aberrant splicing is implicatedin disease, or where an overproduction of a particular splice product isimplicated in disease. Aberrant fusion junctions due to rearrangementsor deletions are also preferred target sites. mRNA transcripts producedvia the process of splicing of two (or more) mRNAs from different genesources are known as “fusion transcripts”. It is also known that intronscan be effectively targeted using antisense compounds targeted to, forexample, DNA or pre-mRNA.

[0082] It is also known in the art that alternative RNA transcripts canbe produced from the same genomic region of DNA. These alternativetranscripts are generally known as “variants”. More specifically,“pre-mRNA variants” are transcripts produced from the same genomic DNAthat differ from other transcripts produced from the same genomic DNA ineither their start or stop position and contain both intronic and exonicsequence.

[0083] Upon excision of one or more exon or intron regions, or portionsthereof during splicing, pre-mRNA variants produce smaller “mRNAvariants”. Consequently, mRNA variants are processed pre-mRNA variantsand each unique pre-mRNA variant must always produce a unique mRNAvariant as a result of splicing. These mRNA variants are also known as“alternative splice variants”. If no splicing of the pre-mRNA variantoccurs then the pre-mRNA variant is identical to the mRNA variant.

[0084] It is also known in the art that variants can be produced throughthe use of alternative signals to start or stop transcription and thatpre-mRNAs and mRNAs can possess more that one start codon or stop codon.Variants that originate from a pre-mRNA or mRNA that use alternativestart codons are known as “alternative start variants” of that pre-mRNAor mRNA. Those transcripts that use an alternative stop codon are knownas “alternative stop variants” of that pre-mRNA or mRNA. One specifictype of alternative stop variant is the “polyA variant” in which themultiple transcripts produced result from the alternative selection ofone of the “polyA stop signals” by the transcription machinery, therebyproducing transcripts that terminate at unique polyA sites. Within thecontext of the invention, the types of variants described herein arealso preferred target nucleic acids.

[0085] The locations on the target nucleic acid to which the preferredantisense compounds hybridize are hereinbelow referred to as “preferredtarget segments.” As used herein the term “preferred target segment” isdefined as at least an 8-nucleobase portion of a target region to whichan active antisense compound is targeted. While not wishing to be boundby theory, it is presently believed that these target segments representportions of the target nucleic acid which are accessible forhybridization.

[0086] While the specific sequences of certain preferred target segmentsare set forth herein, one of skill in the art will recognize that theseserve to illustrate and describe particular embodiments within the scopeof the present invention. Additional preferred target segments may beidentified by one having ordinary skill.

[0087] Target segments 8-80 nucleobases in length comprising a stretchof at least eight (8) consecutive nucleobases selected from within theillustrative preferred target segments are considered to be suitable fortargeting as well.

[0088] Target segments can include DNA or RNA sequences that comprise atleast the 8 consecutive nucleobases from the 5′-terminus of one of theillustrative preferred target segments (the remaining nucleobases beinga consecutive stretch of the same DNA or RNA beginning immediatelyupstream of the 5′-terminus of the target segment and continuing untilthe DNA or RNA contains about 8 to about 80 nucleobases). Similarlypreferred target segments are represented by DNA or RNA sequences thatcomprise at least the 8 consecutive nucleobases from the 3′-terminus ofone of the illustrative preferred target segments (the remainingnucleobases being a consecutive stretch of the same DNA or RNA beginningimmediately downstream of the 3′-terminus of the target segment andcontinuing until the DNA or RNA contains about 8 to about 80nucleobases). One having skill in the art armed with the preferredtarget segments illustrated herein will be able, without undueexperimentation, to identify further preferred target segments.

[0089] Once one or more target regions, segments or sites have beenidentified, antisense compounds are chosen which are sufficientlycomplementary to the target, i.e., hybridize sufficiently well and withsufficient specificity, to give the desired effect.

[0090] D. Screening and Target Validation

[0091] In a further embodiment, the “preferred target segments”identified herein may be employed in a screen for additional compoundsthat modulate the expression of HIF1α or HIF2α. “Modulators” are thosecompounds that decrease or increase the expression of a nucleic acidmolecule encoding HIF1α or HIF2α and which comprise at least an8-nucleobase portion which is complementary to a preferred targetsegment. The screening method comprises the steps of contacting apreferred target segment of a nucleic acid molecule encoding HIF1α orHIF2α with one or more candidate modulators, and selecting for one ormore candidate modulators which decrease or increase the expression of anucleic acid molecule encoding HIF1α or HIF2α. Once it is shown that thecandidate modulator or modulators are capable of modulating (e.g. eitherdecreasing or increasing) the expression of a nucleic acid moleculeencoding HIF1α or HIF2α, the modulator may then be employed in furtherinvestigative studies of the function of HIF1α or HIF2α, or for use as aresearch, diagnostic, or therapeutic agent in accordance with thepresent invention.

[0092] The preferred target segments of the present invention may bealso be combined with their respective complementary antisense compoundsof the present invention to form stabilized double-stranded (duplexed)oligonucleotides.

[0093] Such double stranded oligonucleotide moieties have been shown inthe art to modulate target expression and regulate translation as wellas RNA processsing via an antisense mechanism. Moreover, thedouble-stranded moieties may be subject to chemical modifications (Fireet al., Nature, 1998, 391, 806-811; Timmons and Fire, Nature 1998, 395,854; Timmons et al., Gene, 2001, 263, 103-112; Tabara et al., Science,1998, 282, 430-431; Montgomery et al., Proc. Natl. Acad. Sci. USA, 1998,95, 15502-15507; Tuschl et al., Genes Dev., 1999, 13, 3191-3197;Elbashir et al., Nature, 2001, 411, 494-498; Elbashir et al., Genes Dev.2001, 15, 188-200). For example, such double-stranded moieties have beenshown to inhibit the target by the classical hybridization of antisensestrand of the duplex to the target, thereby triggering enzymaticdegradation of the target (Tijsterman et al., Science, 2002, 295,694-697).

[0094] The compounds of the present invention can also be applied in theareas of drug discovery and target validation. The present inventioncomprehends the use of the compounds and preferred target segmentsidentified herein in drug discovery efforts to elucidate relationshipsthat exist between HIF1α or HIF2α and a disease state, phenotype, orcondition. These methods include detecting or modulating HIF1α or HIF2αcomprising contacting a sample, tissue, cell, or organism with thecompounds of the present invention, measuring the nucleic acid orprotein level of HIF1α or HIF2α and/or a related phenotypic or chemicalendpoint at some time after treatment, and optionally comparing themeasured value to a non-treated sample or sample treated with a furthercompound of the invention. These methods can also be performed inparallel or in combination with other experiments to determine thefunction of unknown genes for the process of target validation or todetermine the validity of a particular gene product as a target fortreatment or prevention of a particular disease, condition, orphenotype.

[0095] E. Kits, Research Reagents, Diagnostics, and Therapeutics

[0096] The compounds of the present invention can be utilized fordiagnostics, therapeutics, prophylaxis and as research reagents andkits. Furthermore, antisense oligonucleotides, which are able to inhibitgene expression with exquisite specificity, are often used by those ofordinary skill to elucidate the function of particular genes or todistinguish between functions of various members of a biologicalpathway.

[0097] For use in kits and diagnostics, the compounds of the presentinvention, either alone or in combination with other compounds ortherapeutics, can be used as tools in differential and/or combinatorialanalyses to elucidate expression patterns of a portion or the entirecomplement of genes expressed within cells and tissues.

[0098] As one nonlimiting example, expression patterns within cells ortissues treated with one or more antisense compounds are compared tocontrol cells or tissues not treated with antisense compounds and thepatterns produced are analyzed for differential levels of geneexpression as they pertain, for example, to disease association,signaling pathway, cellular localization, expression level, size,structure or function of the genes examined. These analyses can beperformed on stimulated or unstimulated cells and in the presence orabsence of other compounds which affect expression patterns.

[0099] Examples of methods of gene expression analysis known in the artinclude DNA arrays or microarrays (Brazma and Vilo, FEBS Lett., 2000,480, 17-24; Celis, et al., FEBS Lett., 2000, 480, 2-16), SAGE (serialanalysis of gene expression) (Madden, et al., Drug Discov. Today, 2000,5, 415-425), READS (restriction enzyme amplification of digested cDNAs)(Prashar and Weissman, Methods Enzymol., 1999, 303, 258-72), TOGA (totalgene expression analysis) (Sutcliffe, et al., Proc. Natl. Acad. Sci.U.S.A., 2000, 97, 1976-81), protein arrays and proteomics (Celis, etal., FEBS Lett., 2000, 480, 2-16; Jungblut, et al., Electrophoresis,1999, 20, 2100-10), expressed sequence tag (EST) sequencing (Celis, etal., FEBS Lett., 2000, 480, 2-16; Larsson, et al., J. Biotechnol., 2000,80, 143-57), subtractive RNA fingerprinting (SuRF) (Fuchs, et al., Anal.Biochem., 2000, 286, 91-98; Larson, et al., Cytometry, 2000, 41,203-208), subtractive cloning, differential display (DD) (Jurecic andBelmont, Curr. Opin. Microbiol., 2000, 3, 316-21), comparative genomichybridization (Carulli, et al., J. Cell Biochem. Suppl., 1998, 31,286-96), FISH (fluorescent in situ hybridization) techniques (Going andGusterson, Eur. J. Cancer, 1999, 35, 1895-904) and mass spectrometrymethods (To, Comb. Chem. High Throughput Screen, 2000, 3, 235-41).

[0100] The compounds of the invention are useful for research anddiagnostics, because these compounds hybridize to nucleic acids encodingHIF1α or HIF2α. For example, oligonucleotides that are shown tohybridize with such efficiency and under such conditions as disclosedherein as to be effective HIF1α or HIF2α inhibitors will also beeffective primers or probes under conditions favoring gene amplificationor detection, respectively. These primers and probes are useful inmethods requiring the specific detection of nucleic acid moleculesencoding HIF1α or HIF2α and in the amplification of said nucleic acidmolecules for detection or for use in further studies of HIF1α or HIF2α.Hybridization of the antisense oligonucleotides, particularly theprimers and probes, of the invention with a nucleic acid encoding HIF1αor HIF2α can be detected by means known in the art. Such means mayinclude conjugation of an enzyme to the oligonucleotide, radiolabellingof the oligonucleotide or any other suitable detection means. Kits usingsuch detection means for detecting the level of HIF1α or HIF2α in asample may also be prepared.

[0101] The specificity and sensitivity of antisense is also harnessed bythose of skill in the art for therapeutic uses. Antisense compounds havebeen employed as therapeutic moieties in the treatment of disease statesin animals, including humans. Antisense oligonucleotide drugs, includingribozymes, have been safely and effectively administered to humans andnumerous clinical trials are presently underway. It is thus establishedthat antisense compounds can be useful therapeutic modalities that canbe configured to be useful in treatment regimes for the treatment ofcells, tissues and animals, especially humans.

[0102] For therapeutics, an animal, preferably a human, suspected ofhaving a disease or disorder which can be treated by modulating theexpression of HIF1α or HIF2α is treated by administering one or moreantisense compounds in accordance with this invention. For example, inone non-limiting embodiment, the methods comprise the step ofadministering to the animal in need of treatment, a therapeuticallyeffective amount of a HIF1α or HIF2α inhibitor. The HIF1α or HIF2αinhibitors of the present invention effectively inhibit the activity ofthe HIF target protein or inhibit the expression of the HIF1α or HIF2αprotein. In one embodiment, the activity or expression of HIF1α or HIF2αin an animal is inhibited by about 10%. Preferably, the activity orexpression of HIF1α or HIF2α in an animal is inhibited by about 30%.More preferably, the activity or expression of HIF1α and/or HIF2α in ananimal is inhibited by 50% or more.

[0103] For example, the reduction of the expression of HIF1α may bemeasured in serum, adipose tissue, liver or any other body fluid, tissueor organ of the animal. Preferably, the cells contained within saidfluids, tissues or organs being analyzed contain a nucleic acid moleculeencoding HIF1α or HIF2α protein and/or the HIF1α or HIF2α proteinitself.

[0104] The compounds of the invention can be utilized in pharmaceuticalcompositions by adding an effective amount of a compound to a suitablepharmaceutically acceptable diluent or carrier. Use of the compounds andmethods of the invention may also be useful prophylactically.

[0105] F. Modifications

[0106] As is known in the art, a nucleoside is a base-sugar combination.The base portion of the nucleoside is normally a heterocyclic base. Thetwo most common classes of such heterocyclic bases are the purines andthe pyrimidines. Nucleotides are nucleosides that further include aphosphate group covalently linked to the sugar portion of thenucleoside. For those nucleosides that include a pentofuranosyl sugar,the phosphate group can be linked to either the 2′, 3′ or 5′ hydroxylmoiety of the sugar. In forming oligonucleotides, the phosphate groupscovalently link adjacent nucleosides to one another to form a linearpolymeric compound. In turn, the respective ends of this linearpolymeric compound can be further joined to form a circular compound,however, linear compounds are generally preferred. In addition, linearcompounds may have internal nucleobase complementarity and may thereforefold in a manner as to produce a fully or partially double-strandedcompound. Within oligonucleotides, the phosphate groups are commonlyreferred to as forming the internucleoside backbone of theoligonucleotide. The normal linkage or backbone of RNA and DNA is a 3′to 5′ phosphodiester linkage.

[0107] Modified Internucleoside Linkages (Backbones)

[0108] Specific examples of preferred antisense compounds useful in thisinvention include oligonucleotides containing modified backbones ornon-natural internucleoside linkages. As defined in this specification,oligonucleotides having modified backbones include those that retain aphosphorus atom in the backbone and those that do not have a phosphorusatom in the backbone. For the purposes of this specification, and assometimes referenced in the art, modified oligonucleotides that do nothave a phosphorus atom in their internucleoside backbone can also beconsidered to be oligonucleosides.

[0109] Preferred modified oligonucleotide backbones containing aphosphorus atom therein include, for example, phosphorothioates, chiralphosphorothioates, phosphorodithioates, phosphotriesters,aminoalkylphosphotriesters, methyl and other alkyl phosphonatesincluding 3′-alkylene phosphonates, 5′-alkylene phosphonates and chiralphosphonates, phosphinates, phosphoramidates including 3′-aminophosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates,thionoalkylphosphonates, thionoalkylphosphotriesters, selenophosphatesand boranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogsof these, and those having inverted polarity wherein one or moreinternucleotide linkages is a 3′ to 3′, 5′ to 5′ or 2′ to 2′ linkage.Preferred oligonucleotides having inverted polarity comprise a single 3′to 3′ linkage at the 3′-most internucleotide linkage i.e. a singleinverted nucleoside residue which may be abasic (the nucleobase ismissing or has a hydroxyl group in place thereof). Various salts, mixedsalts and free acid forms are also included.

[0110] Representative United States patents that teach the preparationof the above phosphorus-containing linkages include, but are not limitedto, U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243;5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717;5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677;5,476,925; 5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253;5,571,799; 5,587,361; 5,194,599; 5,565,555; 5,527,899; 5,721,218;5,672,697 and 5,625,050, certain of which are commonly owned with thisapplication, and each of which is herein incorporated by reference.

[0111] Preferred modified oligonucleotide backbones that do not includea phosphorus atom therein have backbones that are formed by short chainalkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkylor cycloalkyl internucleoside linkages, or one or more short chainheteroatomic or heterocyclic internucleoside linkages. These includethose having morpholino linkages (formed in part from the sugar portionof a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfonebackbones; formacetyl and thioformacetyl backbones; methylene formacetyland thioformacetyl backbones; riboacetyl backbones; alkene containingbackbones; sulfamate backbones; methyleneimino and methylenehydrazinobackbones; sulfonate and sulfonamide backbones; amide backbones; andothers having mixed N, O, S and CH₂ component parts.

[0112] Representative United States patents that teach the preparationof the above oligonucleosides include, but are not limited to, U.S. Pat.Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033;5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967;5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,610,289;5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312;5,633,360; 5,677,437; 5,792,608; 5,646,269 and 5,677,439, certain ofwhich are commonly owned with this application, and each of which isherein incorporated by reference.

[0113] Modified Sugar and Internucleoside Linkages-Mimetics

[0114] In other preferred oligonucleotide mimetics, both the sugar andthe internucleoside linkage (i.e. the backbone), of the nucleotide unitsare replaced with novel groups. The nucleobase units are maintained forhybridization with an appropriate target nucleic acid. One suchcompound, an oligonucleotide mimetic that has been shown to haveexcellent hybridization properties, is referred to as a peptide nucleicacid (PNA). In PNA compounds, the sugar-backbone of an oligonucleotideis replaced with an amide containing backbone, in particular anaminoethylglycine backbone. The nucleobases are retained and are bounddirectly or indirectly to aza nitrogen atoms of the amide portion of thebackbone. Representative United States patents that teach thepreparation of PNA compounds include, but are not limited to, U.S. Pat.Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is hereinincorporated by reference. Further teaching of PNA compounds can befound in Nielsen et al., Science, 1991, 254, 1497-1500.

[0115] Preferred embodiments of the invention are oligonucleotides withphosphorothioate backbones and oligonucleosides with heteroatombackbones, and in particular —CH₂—NH—O—CH₂—, —CH₂—N(CH₃)—O—CH₂— [knownas a methylene (methylimino) or MMI backbone], —CH₂—O—N(CH₃)—CH₂—,—CH₂—N(CH₃)—N(CH₃)—CH₂— and —O—N(CH₃)—CH₂—CH₂— [wherein the nativephosphodiester backbone is represented as —O—P—O—CH₂—] of the abovereferenced U.S. Pat. No. 5,489,677, and the amide backbones of the abovereferenced U.S. Pat. No. 5,602,240. Also preferred are oligonucleotideshaving morpholino backbone structures of the above-referenced U.S. Pat.No. 5,034,506.

[0116] Modified Sugars

[0117] Modified oligonucleotides may also contain one or moresubstituted sugar moieties. Preferred oligonucleotides comprise one ofthe following at the 2′ position: OH; F; O-, S-, or N-alkyl; O-, S-, orN-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl,alkenyl and alkynyl may be substituted or unsubstituted C₁ to C₁₀ alkylor C₂ to C₁₀ alkenyl and alkynyl. Particularly preferred areO[(CH₂)_(n)O]_(m)CH₃, O(CH₂)_(n)OCH₃, O(CH₂)_(n)NH₂, O(CH₂)_(n)CH₃,O(CH₂)_(n)ONH₂, and O(CH₂)_(n)ON[(CH₂)_(n)CH₃]₂, where n and m are from1 to about 10. Other preferred oligonucleotides comprise one of thefollowing at the 2′ position: C₁ to C₁₀ lower alkyl, substituted loweralkyl, alkenyl, alkynyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH,SCH₃, OCN, Cl, Br, CN, CF₃, OCF₃, SOCH₃, SO₂CH₃, ONO₂, NO₂, N₃, NH₂,heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino,substituted silyl, an RNA cleaving group, a reporter group, anintercalator, a group for improving the pharmacokinetic properties of anoligonucleotide, or a group for improving the pharmacodynamic propertiesof an oligonucleotide, and other substituents having similar properties.A preferred modification includes 2′-methoxyethoxy (2′-O—CH₂CH₂OCH₃,also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al., Helv.Chim. Acta, 1995, 78, 486-504) i.e., an alkoxyalkoxy group. A furtherpreferred modification includes 2′-dimethylaminooxyethoxy, i.e., aO(CH₂)₂ON(CH₃)₂ group, also known as 2′-DMAOE, as described in exampleshereinbelow, and 2′-dimethylaminoethoxyethoxy (also known in the art as2′-O-dimethyl-amino-ethoxy-ethyl or 2′-DMAEOE), i.e.,2′-O—CH₂—O—CH₂—N(CH₃)₂, also described in examples hereinbelow.

[0118] Other preferred modifications include 2′-methoxy (2′-O—CH₃),2′-aminopropoxy (2′-OCH₂CH₂CH₂NH₂), 2′-allyl (2′-CH₂—CH═CH₂), 2′-O-allyl(2′-O—CH₂—CH═CH₂) and 2′-fluoro (2′-F). The 2′-modification may be inthe arabino (up) position or ribo (down) position. A preferred2′-arabino modification is 2′-F. Similar modifications may also be madeat other positions on the oligonucleotide, particularly the 3′ positionof the sugar on the 3′ terminal nucleotide or in 2′-5′ linkedoligonucleotides and the 5′ position of 5′ terminal nucleotide.Oligonucleotides may also have. sugar mimetics such as cyclobutylmoieties in place of the pentofuranosyl sugar. Representative UnitedStates patents that teach the preparation of such modified sugarstructures include, but are not limited to, U.S. Pat. Nos. 4,981,957;5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786;5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909;5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633;5,792,747; and 5,700,920, certain of which are commonly owned with theinstant application, and each of which is herein incorporated byreference in its entirety.

[0119] A further preferred modification of the sugar includes LockedNucleic Acids (LNAs) in which the 2′-hydroxyl group is linked to the 3′or 4′ carbon atom of the sugar ring, thereby forming a bicyclic sugarmoiety. The linkage is preferably a methylene (—CH₂—)_(n) group bridgingthe 2′ oxygen atom and the 4′ carbon atom wherein n is 1 or 2. LNAs andpreparation thereof are described in WO 98/39352 and WO 99/14226.

[0120] Natural and Modified Nucleobases

[0121] Oligonucleotides may also include nucleobase (often referred toin the art simply as “base”) modifications or substitutions. As usedherein, “unmodified” or “natural” nucleobases include the purine basesadenine (A) and guanine (G), and the pyrimidine bases thymine (T),cytosine (C) and uracil (U). Modified nucleobases include othersynthetic and natural nucleobases such as 5-methylcytosine (5-me-C),5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine,6-methyl and other alkyl derivatives of adenine and guanine, 2-propyland other alkyl derivatives of adenine and guanine, 2-thiouracil,2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl(—C≡C—CH₃) uracil and cytosine and other alkynyl derivatives ofpyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil(pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl,8-hydroxyl and other 8-substituted adenines and guanines, 5-haloparticularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracilsand cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine,2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and7-deazaadenine and 3-deazaguanine and 3-deazaadenine. Further modifiednucleobases include tricyclic pyrimidines such as phenoxazinecytidine(1H-pyrimido[5,4-b][1,4]benzoxazin-2(3H)-one), phenothiazinecytidine (1H-pyrimido[5,4-b][1,4]benzothiazin-2(3H)-one), G-clamps suchas a substituted phenoxazine cytidine (e.g.9-(2-aminoethoxy)-H-pyrimido[5,4-b][1,4]benzoxazin-2(3H)-one), carbazolecytidine (2H-pyrimido[4,5-b]indol-2-one), pyridoindole cytidine(H-pyrido[3′,2′:4,5]pyrrolo[2,3-d]pyrimidin-2-one). Modified nucleobasesmay also include those in which the purine or pyrimidine base isreplaced with other heterocycles, for example 7-deaza-adenine,7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobasesinclude those disclosed in U.S. Pat. No. 3,687,808, those disclosed inThe Concise Encyclopedia Of Polymer Science And Engineering, pages858-859, Kroschwitz, J. I., ed. John Wiley & Sons, 1990, those disclosedby Englisch et al., Angewandte Chemie, International Edition, 1991, 30,613, and those disclosed by Sanghvi, Y. S., Chapter 15, AntisenseResearch and Applications, pages 289-302, Crooke, S. T. and Lebleu, B.ed., CRC Press, 1993. Certain of these nucleobases are particularlyuseful for increasing the binding affinity of the compounds of theinvention. These include 5-substituted pyrimidines, 6-azapyrimidines andN-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine,5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutionshave been shown to increase nucleic acid duplex stability by 0.6-1.2° C.and are presently preferred base substitutions, even more particularlywhen combined with 2′-O-methoxyethyl sugar modifications.

[0122] Representative United States patents that teach the preparationof certain of the above noted modified nucleobases as well as othermodified nucleobases include, but are not limited to, the above notedU.S. Pat. No. 3,687,808, as well as U.S. Pat. Nos. 4,845,205; 5,130,302;5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255;5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121,5,596,091; 5,614,617; 5,645,985; 5,830,653; 5,763,588; 6,005,096; and5,681,941, certain of which are commonly owned with the instantapplication, and each of which is herein incorporated by reference, andU.S. Pat. No. 5,750,692, which is commonly owned with the instantapplication and also herein incorporated by reference.

[0123] Conjugates

[0124] Another modification of the oligonucleotides of the inventioninvolves chemically linking to the oligonucleotide one or more moietiesor conjugates which enhance the activity, cellular distribution orcellular uptake of the oligonucleotide. These moieties or conjugates caninclude conjugate groups covalently bound to functional groups such asprimary or secondary hydroxyl groups. Conjugate groups of the inventioninclude intercalators, reporter molecules, polyamines, polyamides,polyethylene glycols, polyethers, groups that enhance thepharmacodynamic properties of oligomers, and groups that enhance thepharmacokinetic properties of oligomers. Typical conjugate groupsinclude cholesterols, lipids, phospholipids, biotin, phenazine, folate,phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines,coumarins, and dyes. Groups that enhance the pharmacodynamic properties,in the context of this invention, include groups that improve uptake,enhance resistance to degradation, and/or strengthen sequence-specifichybridization with the target nucleic acid. Groups that enhance thepharmacokinetic properties, in the context of this invention, includegroups that improve uptake, distribution, metabolism or excretion of thecompounds of the present invention. Representative conjugate groups aredisclosed in International Patent Application PCT/US92/09196, filed Oct.23, 1992, and U.S. Pat. No. 6,287,860, the entire disclosure of whichare incorporated herein by reference. Conjugate moieties include but arenot limited to lipid moieties such as a cholesterol moiety, cholic acid,a thioether, e.g., hexyl-S-tritylthiol, a thiocholesterol, an aliphaticchain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g.,di-hexadecyl-rac-glycerol or triethyl-ammonium1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or apolyethylene glycol chain, or adamantane acetic acid, a palmityl moiety,or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety.Oligonucleotides of the invention may also be conjugated to active drugsubstances, for example, aspirin, warfarin, phenylbutazone, ibuprofen,suprofen, fenbufen, ketoprofen, (S)-(+)-pranoprofen, carprofen,dansylsarcosine, 2,3,5-triiodobenzoic acid, flufenamic acid, folinicacid, a benzothiadiazide, chlorothiazide, a diazepine, indomethicin, abarbiturate, a cephalosporin, a sulfa drug, an antidiabetic, anantibacterial or an antibiotic. Oligonucleotide-drug conjugates andtheir preparation are described in U.S. patent application Ser. No.09/334,130 (filed Jun. 15, 1999) which is incorporated herein byreference in its entirety.

[0125] Representative United States patents that teach the preparationof such oligonucleotide conjugates include, but are not limited to, U.S.Pat. Nos. 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541,313;5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,580,731; 5,591,584;5,109,124; 5,118,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439;5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779;4,789,737; 4,824,941; 4,835,263; 4,876,335; 4,904,582; 4,958,013;5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136;5,245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873;5,317,098; 5,371,241, 5,391,723; 5,416,203, 5,451,463; 5,510,475;5,512,667; 5,514,785; 5,565,552; 5,567,810; 5,574,142; 5,585,481;5,587,371; 5,595,726; 5,597,696; 5,599,923; 5,599,928 and 5,688,941,certain of which are commonly owned with the instant application, andeach of which is herein incorporated by reference.

[0126] Chimeric Compounds

[0127] It is not necessary for all positions in a given compound to beuniformly modified, and in fact more than one of the aforementionedmodifications may be incorporated in a single compound or even at asingle nucleoside within an oligonucleotide.

[0128] The present invention also includes antisense compounds which arechimeric compounds. “Chimeric” antisense compounds or “chimeras,” in thecontext of this invention, are antisense compounds, particularlyoligonucleotides, which contain two or more chemically distinct regions,each made up of at least one monomer unit, i.e., a nucleotide in thecase of an oligonucleotide compound. These oligonucleotides typicallycontain at least one region wherein the oligonucleotide is modified soas to confer upon the oligonucleotide increased resistance to nucleasedegradation, increased cellular uptake, increased stability and/orincreased binding affinity for the target nucleic acid. An additionalregion of the oligonucleotide may serve as a substrate for enzymescapable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNAseH is a cellular endonuclease which cleaves the RNA strand of an RNA:DNAduplex. Activation of RNase H, therefore, results in cleavage of the RNAtarget, thereby greatly enhancing the efficiency ofoligonucleotide-mediated inhibition of gene expression. The cleavage ofRNA:RNA hybrids can, in like fashion, be accomplished through theactions of endoribonucleases, such as RNAseL which cleaves both cellularand viral RNA. Cleavage of the RNA target can be routinely detected bygel electrophoresis and, if necessary, associated nucleic acidhybridization techniques known in the art.

[0129] Chimeric antisense compounds of the invention may be formed ascomposite structures of two or more oligonucleotides, modifiedoligonucleotides, oligonucleosides and/or oligonucleotide mimetics asdescribed above. Such compounds have also been referred to in the art ashybrids or gapmers. Representative United States patents that teach thepreparation of such hybrid structures include, but are not limited to,U.S. Pat. Nos. 5,013,830; 5,149,797; 5,220,007; 5,256,775; 5,366,878;5,403,711; 5,491,133; 5,565,350; 5,623,065; 5,652,355; 5,652,356; and5,700,922, certain of which are commonly owned with the instantapplication, and each of which is herein incorporated by reference inits entirety.

[0130] G. Formulations

[0131] The compounds of the invention may also be admixed, encapsulated,conjugated or otherwise associated with other molecules, moleculestructures or mixtures of compounds, as for example, liposomes,receptor-targeted molecules, oral, rectal, topical or otherformulations, for assisting in uptake, distribution and/or absorption.Representative United States patents that teach the preparation of suchuptake, distribution and/or absorption-assisting formulations include,but are not limited to, U.S. Pat. Nos. 5,108,921; 5,354,844; 5,416,016;5,459,127; 5,521,291; 5,543,158; 5,547,932; 5,583,020; 5,591,721;4,426,330; 4,534,899; 5,013,556; 5,108,921; 5,213,804; 5,227,170;5,264,221; 5,356,633; 5,395,619; 5,416,016; 5,417,978; 5,462,854;5,469,854; 5,512,295; 5,527,528; 5,534,259; 5,543,152; 5,556,948;5,580,575; and 5,595,756, each of which is herein incorporated byreference.

[0132] The antisense compounds of the invention encompass anypharmaceutically acceptable salts, esters, or salts of such esters, orany other compound which, upon administration to an animal, including ahuman, is capable of providing (directly or indirectly) the biologicallyactive metabolite or residue thereof. Accordingly, for example, thedisclosure is also drawn to prodrugs and pharmaceutically acceptablesalts of the compounds of the invention, pharmaceutically acceptablesalts of such prodrugs, and other bioequivalents. The term “prodrug”indicates a therapeutic agent that is prepared in an inactive form thatis converted to an active form (i.e., drug) within the body or cellsthereof by the action of endogenous enzymes or other chemicals and/orconditions. In particular, prodrug versions of the oligonucleotides ofthe invention are prepared as SATE [(S-acetyl-2-thioethyl) phosphate]derivatives according to the methods disclosed in WO 93/24510 toGosselin et al., published Dec. 9, 1993 or in WO 94/26764 and U.S. Pat.No. 5,770,713 to Imbach et al.

[0133] The term “pharmaceutically acceptable salts” refers tophysiologically and pharmaceutically acceptable salts of the compoundsof the invention: i.e., salts that retain the desired biologicalactivity of the parent compound and do not impart undesiredtoxicological effects thereto. For oligonucleotides, preferred examplesof pharmaceutically acceptable salts and their uses are furtherdescribed in U.S. Pat. No. 6,287,860, which is incorporated herein inits entirety.

[0134] The present invention also includes pharmaceutical compositionsand formulations which include the antisense compounds of the invention.The pharmaceutical compositions of the present invention may beadministered in a number of ways depending upon whether local orsystemic treatment is desired and upon the area to be treated.Administration may be topical (including ophthalmic and to mucousmembranes including vaginal and rectal delivery), pulmonary, e.g., byinhalation or insufflation of powders or aerosols, including bynebulizer; intratracheal, intranasal, epidermal and transdermal), oralor parenteral. Parenteral administration, includes intravenous,intraarterial, subcutaneous, intraperitoneal or intramuscular injectionor infusion; or intracranial, e.g., intrathecal or intraventricular,administration. Oligonucleotides with at least one 2′-O-methoxyethylmodification are believed to be particularly useful for oraladministration. Pharmaceutical compositions and formulations for topicaladministration may include transdermal patches, ointments, lotions,creams, gels, drops, suppositories, sprays, liquids and powders.Conventional pharmaceutical carriers, aqueous, powder or oily bases,thickeners and the like may be necessary or desirable. Coated condoms,gloves and the like may also be useful.

[0135] The pharmaceutical formulations of the present invention, whichmay conveniently be presented in unit dosage form, may be preparedaccording to conventional techniques well known in the pharmaceuticalindustry. Such techniques include the step of bringing into associationthe active ingredients with the pharmaceutical carrier(s) orexcipient(s). In general, the formulations are prepared by uniformly andintimately bringing into association the active ingredients with liquidcarriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product.

[0136] The compositions of the present invention may be formulated intoany of many possible dosage forms such as, but not limited to, tablets,capsules, gel capsules, liquid syrups, soft gels, suppositories, andenemas. The compositions of the present invention may also be formulatedas suspensions in aqueous, non-aqueous or mixed media. Aqueoussuspensions may further contain substances which increase the viscosityof the suspension including, for example, sodium carboxymethylcellulose,sorbitol and/or dextran. The suspension may also contain stabilizers.

[0137] Pharmaceutical compositions of the present invention include, butare not limited to, solutions, emulsions, foams and liposome-containingformulations. The pharmaceutical compositions and formulations of thepresent invention may comprise one or more penetration enhancers,carriers, excipients or other active or inactive ingredients.

[0138] Emulsions are typically heterogenous systems of one liquiddispersed in another in the form of droplets usually exceeding 0.1 μm indiameter. Emulsions may contain additional components in addition to thedispersed phases, and the active drug which may be present as a solutionin either the aqueous phase, oily phase or itself as a separate phase.Microemulsions are included as an embodiment of the present invention.Emulsions and their uses are well known in the art and are furtherdescribed in U.S. Pat. No. 6,287,860, which is incorporated herein inits entirety.

[0139] Formulations of the present invention include liposomalformulations. As used in the present invention, the term “liposome”means a vesicle composed of amphiphilic lipids arranged in a sphericalbilayer or bilayers. Liposomes are unilamellar or multilamellar vesicleswhich have a membrane formed from a lipophilic material and an aqueousinterior that contains the composition to be delivered. Cationicliposomes are positively charged liposomes which are believed tointeract with negatively charged DNA molecules to form a stable complex.Liposomes that are pH-sensitive or negatively-charged are believed toentrap DNA rather than complex with it. Both cationic and noncationicliposomes have been used to deliver DNA to cells.

[0140] Liposomes also include “sterically stabilized” liposomes, a termwhich, as used herein, refers to liposomes comprising one or morespecialized lipids that, when incorporated into liposomes, result inenhanced circulation lifetimes relative to liposomes lacking suchspecialized lipids. Examples of sterically stabilized liposomes arethose in which part of the vesicle-forming lipid portion of the liposomecomprises one or more glycolipids or is derivatized with one or morehydrophilic polymers, such as a polyethylene glycol (PEG) moiety.Liposomes and their uses are further described in U.S. Pat. No.6,287,860, which is incorporated herein in its entirety.

[0141] The pharmaceutical formulations and compositions of the presentinvention may also include surfactants. The use of surfactants in drugproducts, formulations and in emulsions is well known in the art.Surfactants and their uses are further described in U.S. Pat. No.6,287,860, which is incorporated herein in its entirety.

[0142] In one embodiment, the present invention employs variouspenetration enhancers to effect the efficient delivery of nucleic acids,particularly oligonucleotides. In addition to aiding the diffusion ofnon-lipophilic drugs across cell membranes, penetration enhancers alsoenhance the permeability of lipophilic drugs. Penetration enhancers maybe classified as belonging to one of five broad categories, i.e.,surfactants, fatty acids, bile salts, chelating agents, andnon-chelating non-surfactants. Penetration enhancers and their uses arefurther described in U.S. Pat. No. 6,287,860, which is incorporatedherein in its entirety.

[0143] One of skill in the art will recognize that formulations areroutinely designed according to their intended use, i.e. route ofadministration.

[0144] Preferred formulations for topical administration include thosein which the oligonucleotides of the invention are in admixture with atopical delivery agent such as lipids, liposomes, fatty acids, fattyacid esters, steroids, chelating agents and surfactants. Preferredlipids and liposomes include neutral (e.g. dioleoylphosphatidyl DOPEethanolamine, dimyristoylphosphatidyl choline DMPC,distearolyphosphatidyl choline) negative (e.g. dimyristoylphosphatidylglycerol DMPG) and cationic (e.g. dioleoyltetramethylaminopropyl DOTAPand dioleoylphosphatidyl ethanolamine DOTMA).

[0145] For topical or other administration, oligonucleotides of theinvention may be encapsulated within liposomes or may form complexesthereto, in particular to cationic liposomes. Alternatively,oligonucleotides may be complexed to lipids, in particular to cationiclipids. Preferred fatty acids and esters, pharmaceutically acceptablesalts thereof, and their uses are further described in U.S. Pat. No.6,287,860, which is incorporated herein in its entirety. Topicalformulations are described in detail in U.S. patent application Ser. No.09/315,298 filed on May 20, 1999, which is incorporated herein byreference in its entirety.

[0146] Compositions and formulations for oral administration includepowders or granules, microparticulates, nanoparticulates, suspensions orsolutions in water or non-aqueous media, capsules, gel capsules,sachets, tablets or minitablets. Thickeners, flavoring agents, diluents,emulsifiers, dispersing aids or binders may be desirable. Preferred oralformulations are those in which oligonucleotides of the invention areadministered in conjunction with one or more penetration enhancerssurfactants and chelators. Preferred surfactants include fatty acidsand/or esters or salts thereof, bile acids and/or salts thereof.Preferred bile acids/salts and fatty acids and their uses are furtherdescribed in U.S. Pat. No. 6,287,860, which is incorporated herein inits entirety. Also preferred are combinations of penetration enhancers,for example, fatty acids/salts in combination with bile acids/salts. Aparticularly preferred combination is the sodium salt of lauric acid,capric acid and UDCA. Further penetration enhancers includepolyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether.Oligonucleotides of the invention may be delivered orally, in granularform including sprayed dried particles, or complexed to form micro ornanoparticles. Oligonucleotide complexing agents and their uses arefurther described in U.S. Pat. No. 6,287,860, which is incorporatedherein in its entirety. Oral formulations for oligonucleotides and theirpreparation are described in detail in U.S. application Ser. No.09/108,673 (filed Jul. 1, 1998), Ser. No. 09/315,298 (filed May 20,1999) and Ser. No. 10/071,822, filed Feb. 8, 2002, each of which isincorporated herein by reference in their entirety.

[0147] Compositions and formulations for parenteral, intrathecal orintraventricular administration may include sterile aqueous solutionswhich may also contain buffers, diluents and other suitable additivessuch as, but not limited to, penetration enhancers, carrier compoundsand other pharmaceutically acceptable carriers or excipients.

[0148] Certain embodiments of the invention provide pharmaceuticalcompositions containing one or more oligomeric compounds and one or moreother chemotherapeutic agents which function by a non-antisensemechanism. Examples of such chemotherapeutic agents include but are notlimited to cancer chemotherapeutic drugs such as daunorubicin,daunomycin, dactinomycin, doxorubicin, epirubicin, idarubicin,esorubicin, bleomycin, mafosfamide, ifosfamide, cytosine ara-binoside,bis-chloroethylnitrosurea, busulfan, mitomycin C, actinomycin D,mithramycin, prednisone, hydroxyprogesterone, testosterone, tamoxifen,dacarbazine, procarbazine, hexamethylmelamine, pentamethylmelamine,mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea,nitrogen mustards, melphalan, cyclophosphamide, 6-mercaptopurine,6-thioguanine, cytarabine, 5-azacytidine, hydroxyurea, deoxyco-formycin,4-hydroxyperoxycyclophosphoramide, 5-fluorouracil (5-FU),5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, taxol,vincristine, vinblastine, etoposide (VP-16), trimetrexate, irinotecan,topotecan, gemcitabine, teni-poside, cisplatin and diethylstilbestrol(DES). When used with the compounds of the invention, suchchemotherapeutic agents may be used individually (e.g., 5-FU andoligonucleotide), sequentially (e.g., 5-FU and oligonucleotide for aperiod of time followed by MTX and oligonucleotide), or in combinationwith one or more other such chemotherapeutic agents (e.g., 5-FU, MTX andoligonucleotide, or 5-FU, radiotherapy and oligonucleotide).Anti-inflammatory drugs, including but not limited to nonsteroidalanti-inflammatory drugs and corticosteroids, and antiviral drugs,including but not limited to ribivirin, vidarabine, acyclovir andganciclovir, may also be combined in compositions of the invention.Combinations of antisense compounds and other non-antisense drugs arealso within the scope of this invention. Two or more combined compoundsmay be used together or sequentially.

[0149] In another related embodiment, compositions of the invention maycontain one or more antisense compounds, particularly oligonucleotides,targeted to a first nucleic acid and one or more additional antisensecompounds targeted to a second nucleic acid target. Alternatively,compositions of the invention may contain two or more antisensecompounds targeted to different regions of the same nucleic acid target.Numerous examples of antisense compounds are known in the art. Two ormore combined compounds may be used together or sequentially.

[0150] H. Dosing

[0151] The formulation of therapeutic compositions and their subsequentadministration (dosing) is believed to be within the skill of those inthe art. Dosing is dependent on severity and responsiveness of thedisease state to be treated, with the course of treatment lasting fromseveral days to several months, or until a cure is effected or adiminution of the disease state is achieved. Optimal dosing schedulescan be calculated from measurements of drug accumulation in the body ofthe patient. Persons of ordinary skill can easily determine optimumdosages, dosing methodologies and repetition rates. Optimum dosages mayvary depending on the relative potency of individual oligonucleotides,and can generally be estimated based on EC₅₀s found to be effective inin vitro and in vivo animal models. In general, dosage is from 0.01 ugto 100 g per kg of body weight, and may be given once or more daily,weekly, monthly or yearly, or even once every 2 to 20 years. Persons ofordinary skill in the art can easily estimate repetition rates fordosing based on measured residence times and concentrations of the drugin bodily fluids or tissues. Following successful treatment, it may bedesirable to have the patient undergo maintenance therapy to prevent therecurrence of the disease state, wherein the oligonucleotide isadministered in maintenance doses, ranging from 0.01 ug to 100 g per kgof body weight, once or more daily, to once every 20 years.

[0152] While the present invention has been described with specificityin accordance with certain of its preferred embodiments, the followingexamples serve only to illustrate the invention and are not intended tolimit the same.

EXAMPLES Example 1 Synthesis of Nucleoside Phosphoramidites

[0153] The following compounds, including amidites and theirintermediates were prepared as described in U.S. Pat. No. 6,426,220 andpublished PCT WO 02/36743; 5′-O-Dimethoxytrityl-thymidine intermediatefor 5-methyl dC amidite, 5′-O-Dimethoxytrityl-2′-deoxy-5-methylcytidineintermediate for 5-methyl-dC amidite,5′-O-Dimethoxytrityl-2′-deoxy-N4-benzoyl-5-methylcytidine penultimateintermediate for 5-methyl dC amidite,[5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-deoxy-N⁴-benzoyl-5-methylcytidin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite(5-methyl dC amidite), 2′-Fluorodeoxyadenosine, 2′-Fluorodeoxyguanosine,2′-Fluorouridine, 2′-Fluorodeoxycytidine, 2′-O-(2-Methoxyethyl) modifiedamidites, 2′-O-(2-methoxyethyl)-5-methyluridine intermediate,5′-O-DMT-2′-O-(2-methoxyethyl)-5-methyluridine penultimate intermediate,[5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-5-methyluridin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite(MOE T amidite),5′-O-Dimethoxytrityl-2′-O-(2-methoxyethyl)-5-methylcytidineintermediate,5′-O-dimethoxytrityl-2′-O-(2-methoxyethyl)-N⁴-benzoyl-5-methyl-cytidinepenultimate intermediate,[5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N⁴-benzoyl-5-methylcytidin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite(MOE 5-Me-C amidite),[5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N⁶-benzoyladenosin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite(MOE A amdite),[5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N⁴-isobutyrylguanosin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite(MOE G amidite), 2′-O-(Aminooxyethyl) nucleoside amidites and2′-O-(dimethylaminooxyethyl) nucleoside amidites,2′-(Dimethylaminooxyethoxy) nucleoside amidites,5′-O-tert-Butyldiphenylsilyl-O²-2′-anhydro-5-methyluridine ,5′-O-tert-Butyldiphenylsilyl-2′-O-(2-hydroxyethyl)-5-methyluridine,2′-O-([2-phthalimidoxy)ethyl]-5′-t-butyldiphenylsilyl-5-methyluridine,5′-O-tert-butyldiphenylsilyl-2′-O-[(2-formadoximinooxy)ethyl]-5-methyluridine,5′-O-tert-Butyldiphenylsilyl-2′-O-[N,N-dimethylaminooxyethyl]-5-methyluridine,2′-O-(dimethylaminooxyethyl)-5-methyluridine,5′-O-DMT-2′-O-(dimethylaminooxyethyl)-5-methyluridine,5′-O-DMT-2′-O-(2-N,N-dimethylaminooxyethyl)-5-methyluridine-3′-[(2-cyanoethyl)-N,N-diisopropylphosphoramidite],2′-(Aminooxyethoxy) nucleoside amidites,N2-isobutyryl-6-O-diphenylcarbamoyl-2′-O-(2-ethylacetyl)-5′-O-(4,4′-dimethoxytrityl)guanosine-3′-[(2-cyanoethyl)-N,N-diisopropylphosphoramidite],2′-dimethylaminoethoxyethoxy (2′-DMAEOE) nucleoside amidites,2′-O-[2(2-N,N-dimethylaminoethoxy)ethyl]-5-methyl uridine,5′-O-dimethoxytrityl-2′-O-[2(2-N,N-dimethylaminoethoxy)-ethyl)]-5-methyluridine and5′-O-Dimethoxytrityl-2′-O-[2(2-N,N-dimethylaminoethoxy)-ethyl)]-5-methyluridine-3′-O-(cyanoethyl-N,N-diisopropyl)phosphoramidite.

Example 2 Oligonucleotide and Oligonucleoside Synthesis

[0154] The antisense compounds used in accordance with this inventionmay be conveniently and routinely made through the well-known techniqueof solid phase synthesis. Equipment for such synthesis is sold byseveral vendors including, for example, Applied Biosystems (Foster City,Calif.). Any other means for such synthesis known in the art mayadditionally or alternatively be employed. It is well known to usesimilar techniques to prepare oligonucleotides such as thephosphorothioates and alkylated derivatives.

[0155] Oligonucleotides:

[0156] Unsubstituted and substituted phosphodiester (P═O)oligonucleotides are synthesized on an automated DNA synthesizer(Applied Biosystems model 394) using standard phosphoramidite chemistrywith oxidation by iodine.

[0157] Phosphorothioates (P═S) are synthesized similar to phosphodiesteroligonucleotides with the following exceptions: thiation was effected byutilizing a 10% w/v solution of 3,H-1,2-benzodithiole-3-one 1,1-dioxidein acetonitrile for the oxidation of the phosphite linkages. Thethiation reaction step time was increased to 180 sec and preceded by thenormal capping step. After cleavage from the CPG column and deblockingin concentrated ammonium hydroxide at 55° C. (12-16 hr), theoligonucleotides were recovered by precipitating with >3 volumes ofethanol from a 1 M NH₄OAc solution. Phosphinate oligonucleotides areprepared as described in U.S. Pat. No. 5,508,270, herein incorporated byreference.

[0158] Alkyl phosphonate oligonucleotides are prepared as described inU.S. Pat. No. 4,469,863, herein incorporated by reference.

[0159] 3′-Deoxy-3′-methylene phosphonate oligonucleotides are preparedas described in U.S. Pat. Nos. 5,610,289 or 5,625,050, hereinincorporated by reference.

[0160] Phosphoramidite oligonucleotides are prepared as described inU.S. Pat. No. 5,256,775 or U.S. Pat. No. 5,366,878, herein incorporatedby reference.

[0161] Alkylphosphonothioate oligonucleotides are prepared as describedin published PCT applications PCT/US94/00902 and PCT/US93/06976(published as WO 94/17093 and WO 94/02499, respectively), hereinincorporated by reference.

[0162] 3′-Deoxy-3′-amino phosphoramidate oligonucleotides are preparedas described in U.S. Pat. No. 5,476,925, herein incorporated byreference.

[0163] Phosphotriester oligonucleotides are prepared as described inU.S. Pat. No. 5,023,243, herein incorporated by reference.

[0164] Borano phosphate oligonucleotides are prepared as described inU.S. Pat. Nos. 5,130,302 and 5,177,198, both herein incorporated byreference.

[0165] Oligonucleosides:

[0166] Methylenemethylimino linked oligonucleosides, also identified asMMI linked oligonucleosides, methylenedimethylhydrazo linkedoligonucleosides, also identified as MDH linked oligonucleosides, andmethylenecarbonylamino linked oligonucleosides, also identified asamide-3 linked oligonucleosides, and methyleneaminocarbonyl linkedoligonucleosides, also identified as amide-4 linked oligonucleosides, aswell as mixed backbone compounds having, for instance, alternating MMIand P═O or P═S linkages are prepared as described in U.S. Pat. Nos.5,378,825, 5,386,023, 5,489,677, 5,602,240 and 5,610,289, all of whichare herein incorporated by reference.

[0167] Formacetal and thioformacetal linked oligonucleosides areprepared as described in U.S. Pat. Nos. 5,264,562 and 5,264,564, hereinincorporated by reference.

[0168] Ethylene oxide linked oligonucleosides are prepared as describedin U.S. Pat. No. 5,223,618, herein incorporated by reference.

Example 3 RNA Synthesis

[0169] In general, RNA synthesis chemistry is based on the selectiveincorporation of various protecting groups at strategic intermediaryreactions. Although one of ordinary skill in the art will understand theuse of protecting groups in organic synthesis, a useful class ofprotecting groups includes silyl ethers. In particular bulky silylethers are used to protect the 5′-hydroxyl in combination with anacid-labile orthoester protecting group on the 2′-hydroxyl. This set ofprotecting groups is then used with standard solid-phase synthesistechnology. It is important to lastly remove the acid labile orthoesterprotecting group after all other synthetic steps. Moreover, the earlyuse of the silyl protecting groups during synthesis ensures facileremoval when desired, without undesired deprotection of 2′ hydroxyl.

[0170] Following this procedure for the sequential protection of the5′-hydroxyl in combination with protection of the 2′-hydroxyl byprotecting groups that are differentially removed and are differentiallychemically labile, RNA oligonucleotides were synthesized.

[0171] RNA oligonucleotides are synthesized in a stepwise fashion. Eachnucleotide is added sequentially (3′- to 5′-direction) to a solidsupport-bound oligonucleotide. The first nucleoside at the 3′-end of thechain is covalently attached to a solid support. The nucleotideprecursor, a ribonucleoside phosphoramidite, and activator are added,coupling the second base onto the 5′-end of the first nucleoside. Thesupport is washed and any unreacted 5′-hydroxyl groups are capped withacetic anhydride to yield 5′-acetyl moieties. The linkage is thenoxidized to the more stable and ultimately desired P(V) linkage. At theend of the nucleotide addition cycle, the 5′-silyl group is cleaved withfluoride. The cycle is repeated for each subsequent nucleotide.

[0172] Following synthesis, the methyl protecting groups on thephosphates are cleaved in 30 minutes utilizing 1 Mdisodium-2-carbamoyl-2-cyanoethylene-1,1-dithiolate trihydrate (S₂Na₂)in DMF. The deprotection solution is washed from the solid support-boundoligonucleotide using water. The support is then treated with 40%methylamine in water for 10 minutes at 55° C. This releases the RNAoligonucleotides into solution, deprotects the exocyclic amines, andmodifies the 2′-groups. The oligonucleotides can be analyzed by anionexchange HPLC at this stage.

[0173] The 2′-orthoester groups are the last protecting groups to beremoved. The ethylene glycol monoacetate orthoester protecting groupdeveloped by Dharmacon Research, Inc. (Lafayette, Colo.), is one exampleof a useful orthoester protecting group which, has the followingimportant properties. It is stable to the conditions of nucleosidephosphoramidite synthesis and oligonucleotide synthesis. However, afteroligonucleotide synthesis the oligonucleotide is treated withmethylamine which not only cleaves the oligonucleotide from the solidsupport but also removes the acetyl groups from the orthoesters. Theresulting 2-ethyl-hydroxyl substituents on the orthoester are lesselectron withdrawing than the acetylated precursor. As a result, themodified orthoester becomes more labile to acid-catalyzed hydrolysis.Specifically, the rate of cleavage is approximately 10 times fasterafter the acetyl groups are removed. Therefore, this orthoesterpossesses sufficient stability in order to be compatible witholigonucleotide synthesis and yet, when subsequently modified, permitsdeprotection to be carried out under relatively mild aqueous conditionscompatible with the final RNA oligonucleotide product.

[0174] Additionally, methods of RNA synthesis are well known in the art(Scaringe, S. A. Ph.D. Thesis, University of Colorado, 1996; Scaringe,S. A., et al., J. Am. Chem. Soc., 1998, 120, 11820-11821; Matteucci, M.D. and Caruthers, M. H. J. Am. Chem. Soc., 1981, 103, 3185-3191;Beaucage, S. L. and Caruthers, M. H. Tetrahedron Lett., 1981, 22,1859-1862; Dahl, B. J., et al., Acta Chem. Scand., 1990, 44, 639-641;Reddy, M. P., et al., Tetrahedrom Lett., 1994, 25, 4311-4314; Wincott,F. et al., Nucleic Acids Res., 1995, 23, 2677-2684; Griffin, B. E., etal., Tetrahedron, 1967, 23, 2301-2313; Griffin, B. E., et al.,Tetrahedron, 1967, 23, 2315-2331).

[0175] RNA antisense compounds (RNA oligonucleotides) of the presentinvention can be synthesized by the methods herein or purchased fromDharmacon Research, Inc (Lafayette, Colo.). Once synthesized,complementary RNA antisense compounds can then be annealed by methodsknown in the art to form double stranded (duplexed) antisense compounds.For example, duplexes can be formed by combining 30 μl of each of thecomplementary strands of RNA oligonucleotides (50 uM RNA oligonucleotidesolution) and 15 μl of 5× annealing buffer (100 mM potassium acetate, 30mM HEPES-KOH pH 7.4, 2 mM magnesium acetate) followed by heating for 1minute at 90° C., then 1 hour at 37° C. The resulting duplexed antisensecompounds can be used in kits, assays, screens, or other methods toinvestigate the role of a target nucleic acid.

Example 4 Synthesis of Chimeric Oligonucleotides

[0176] Chimeric oligonucleotides, oligonucleosides or mixedoligonucleotides/oligonucleosides of the invention can be of severaldifferent types. These include a first type wherein the “gap” segment oflinked nucleosides is positioned between 5′ and 3′ “wing” segments oflinked nucleosides and a second “open end” type wherein the “gap”segment is located at either the 3′ or the 5′ terminus of the oligomericcompound. Oligonucleotides of the first type are also known in the artas “gapmers” or gapped oligonucleotides. Oligonucleotides of the secondtype are also known in the art as “hemimers” or “wingmers”.

[0177] [2′-O-Me]--[2′-deoxy]--[2′-O-Me] Chimeric PhosphorothioateOligonucleotides

[0178] Chimeric oligonucleotides having 2′-O-alkyl phosphorothioate and2′-deoxy phosphorothioate oligonucleotide segments are synthesized usingan Applied Biosystems automated DNA synthesizer Model 394, as above.Oligonucleotides are synthesized using the automated synthesizer and2′-deoxy-5′-dimethoxytrityl-3′-O-phosphoramidite for the DNA portion and5′-dimethoxytrityl-2′-O-methyl-3′-O-phosphoramidite for 5′ and 3′ wings.The standard synthesis cycle is modified by incorporating coupling stepswith increased reaction times for the5′-dimethoxytrityl-2′-O-methyl-3′-O-phosphoramidite. The fully protectedoligonucleotide is cleaved from the support and deprotected inconcentrated ammonia (NH₄OH) for 12-16 hr at 55° C. The deprotectedoligo is then recovered by an appropriate method (precipitation, columnchromatography, volume reduced in vacuo and analyzedspetrophotometrically for yield and for purity by capillaryelectrophoresis and by mass spectrometry.

[0179] [2′-O-(2-Methoxyethyl)]--[2′-deoxy]--[2′-O-(Methoxyethyl)]Chimeric Phosphorothioate Oligonucleotides

[0180] [2′-O-(2-methoxyethyl)]--[2′-deoxy]--[-2′-O-(methoxyethyl)]chimeric phosphorothioate oligonucleotides were prepared as per theprocedure above for the 2′-O-methyl chimeric oligonucleotide, with thesubstitution of 2′-O-(methoxyethyl) amidites for the 2′-O-methylamidites.

[0181] [2′-O-(2-Methoxyethyl)Phosphodiester]--[2′-deoxyPhosphorothioate]--[2′-O-(2-Methoxyethyl) Phosphodiester] ChimericOligonucleotides

[0182] [2′-O-(2-methoxyethyl phosphodiester]--[2′-deoxyphosphorothioate]--[2′-O-(methoxyethyl) phosphodiester] chimericoligonucleotides are prepared as per the above procedure for the2′-O-methyl chimeric oligonucleotide with the substitution of2′-O-(methoxyethyl) amidites for the 2′-O-methyl amidites, oxidationwith iodine to generate the phosphodiester internucleotide linkageswithin the wing portions of the chimeric structures and sulfurizationutilizing 3,H-1,2 benzodithiole-3-one 1,1 dioxide (Beaucage Reagent) togenerate the phosphorothioate internucleotide linkages for the centergap.

[0183] Other chimeric oligonucleotides, chimeric oligonucleosides andmixed chimeric oligonucleotides/oligonucleosides are synthesizedaccording to U.S. Pat. No. 5,623,065, herein incorporated by reference.

Example 5 D Sign and Screening of Duplexed Antisense Compounds TargetingHIF1α or HIF2α

[0184] In accordance with the present invention, a series of nucleicacid duplexes comprising the antisense compounds of the presentinvention and their complements can be designed to target HIF1α orHIF2α. The nucleobase sequence of the antisense strand of the duplexpreferably comprises at least a portion of an oligonucleotide in Tables1, 3, 4, 5, 6, 13, or 14. The ends of the strands may be modified by theaddition of one or more natural or modified nucleobases to form anoverhang. The sense strand of the dsRNA is then designed and synthesizedas the complement of the antisense strand and may also containmodifications or additions to either terminus. For example, in oneembodiment, both strands of the dsRNA duplex would be complementary overthe central nucleobases, each having overhangs at one or both termini.

[0185] For example, a duplex comprising an antisense strand having thesequence CGAGAGGCGGACGGGACCG and having a two-nucleobase overhang ofdeoxythymidine(dT) would have the following structure:  cgagaggcggacgggaccgTT Antisense Strand   |||||||||||||||||||TTgctctccgcctgccctggc Complement

[0186] As another example, a duplex comprising an antisense strandhaving the sequence CGAGAGGCGGACGGGACCG and having no overhangs wouldhave the following structure: cgagaggcggacgggaccg Antisense Strand||||||||||||||||||| gctctccgcctgccctggc Complement

[0187] RNA strands of the duplex can be synthesized by methods disclosedherein or purchased from Dharmacon Research Inc., (Lafayette, Colo.).Once synthesized, the complementary strands are annealed. The singlestrands are aliquoted and diluted to a concentration of 50 uM. Oncediluted, 30 uL of each strand is combined with 15uL of a 5× solution ofannealing buffer. The final concentration of said buffer is 100 mMpotassium acetate, 30 mM HEPES-KOH pH 7.4, and 2 mM magnesium acetate.The final volume is 75 uL. This solution is incubated for 1 minute at90° C. and then centrifuged for 15 seconds. The tube is allowed to sitfor 1 hour at 37° C. at which time the dsRNA duplexes are used inexperimentation. The final concentration of the dsRNA duplex is 20 uM.This solution can be stored frozen (−20° C.) and freeze-thawed up to 5times.

[0188] Once prepared, the duplexed antisense compounds are evaluated fortheir ability to modulate HIF1• or HIF2• expression.

[0189] When cells reached 80% confluency, they are treated with duplexedantisense compounds of the invention. For cells grown in 96-well plates,wells are washed once with 200 •L OPTI-MEM-1 reduced-serum medium (GibcoBRL) and then treated with 130 •L of OPTI-MEM-1 containing 12 •g/mLLIPOFECTIN (Gibco BRL) and the desired duplex antisense compound at afinal concentration of 200 M. After 5 hours of treatment, the medium isreplaced with fresh medium. Cells are harvested 16 hours aftertreatment, at which time RNA is isolated and target reduction measuredby RT-PCR.

Example 6 Oligonucleotide Isolation

[0190] After cleavage from the controlled pore glass solid support anddeblocking in concentrated ammonium hydroxide at 55° C. for 12-16 hours,the oligonucleotides or oligonucleosides are recovered by precipitationout of 1 M NH₄OAc with >3 volumes of ethanol. Synthesizedoligonucleotides were analyzed by electrospray mass spectroscopy(molecular weight determination) and by capillary gel electrophoresisand judged to be at least 70% full length material. The relative amountsof phosphorothioate and phosphodiester linkages obtained in thesynthesis was determined by the ratio of correct molecular weightrelative to the −16 amu product (+/−32 +/−48). For some studiesoligonucleotides were purified by HPLC, as described by Chiang et al.,J. Biol. Chem. 1991, 266, 18162-18171. Results obtained withHPLC-purified material were similar to those obtained with non-HPLCpurified material.

Example 7 Oligonucleotide Synthesis—96 Well Plate Format

[0191] Oligonucleotides were synthesized via solid phase P(III)phosphoramidite chemistry on an automated synthesizer capable ofassembling 96 sequences simultaneously in a 96-well format.Phosphodiester internucleotide linkages were afforded by oxidation withaqueous iodine. Phosphorothioate internucleotide linkages were generatedby sulfurization utilizing 3,H-1,2 benzodithiole-3-one 1,1 dioxide(Beaucage Reagent) in anhydrous acetonitrile. Standard base-protectedbeta-cyanoethyl-diiso-propyl phosphoramidites were purchased fromcommercial vendors (e.g. PE-Applied Biosystems, Foster City, Calif., orPharmacia, Piscataway, N.J.). Non-standard nucleosides are synthesizedas per standard or patented methods. They are utilized as base protectedbeta-cyanoethyldiisopropyl phosphoramidites.

[0192] Oligonucleotides were cleaved from support and deprotected withconcentrated NH₄OH at elevated temperature (55-60° C.) for 12-16 hoursand the released product then dried in vacuo. The dried product was thenre-suspended in sterile water to afford a master plate from which allanalytical and test plate samples are then diluted utilizing roboticpipettors.

Example 8 Oligonucleotide Analysis—96-Well Plate Format

[0193] The concentration of oligonucleotide in each well was assessed bydilution of samples and UV absorption spectroscopy. The full-lengthintegrity of the individual products was evaluated by capillaryelectrophoresis (CE) in either the 96-well format (Beckman P/ACE™ MDQ)or, for individually prepared samples, on a commercial CE apparatus(e.g., Beckman P/ACE™ 5000, ABI 270). Base and backbone composition wasconfirmed by mass analysis of the compounds utilizing electrospray-massspectroscopy. All assay test plates were diluted from the master plateusing single and multi-channel robotic pipettors. Plates were judged tobe acceptable if at least 85% of the compounds on the plate were atleast 85% full length.

Example 9 Cell Culture and Oligonucleotide Treatment

[0194] The effect of antisense compounds on target nucleic acidexpression can be tested in any of a variety of cell types provided thatthe target nucleic acid is present at measurable levels. This can beroutinely determined using, for example, PCR or Northern blot analysis.The following cell types are provided for illustrative purposes, butother cell types can be routinely used, provided that the target isexpressed in the cell type chosen. This can be readily determined bymethods routine in the art, for example Northern blot analysis,ribonuclease protection assays, or RT-PCR.

[0195] T-24 Cells:

[0196] The human transitional cell bladder carcinoma cell line T-24 wasobtained from the American Type Culture Collection (ATCC) (Manassas,Va.). T-24 cells were routinely cultured in complete McCoy's 5A basalmedia (Invitrogen Corporation, Carlsbad, Calif.) supplemented with 10%fetal calf serum (Invitrogen Corporation, Carlsbad, Calif.), penicillin100 units per mL, and streptomycin 100 micrograms per mL (InvitrogenCorporation, Carlsbad, Calif.). Cells were routinely passaged bytrypsinization and dilution when they reached 90% confluence. Cells wereseeded into 96-well plates (Falcon-Primaria #353872) at a density of7000 cells/well for use in RT-PCR analysis.

[0197] For Northern blotting or other analysis, cells may be seeded onto100 mm or other standard tissue culture plates and treated similarly,using appropriate volumes of medium and oligonucleotide.

[0198] A549 Cells:

[0199] The human lung carcinoma cell line A549 was obtained from theAmerican Type Culture Collection (ATCC) (Manassas, Va.). A549 cells wereroutinely cultured in DMEM basal media (Invitrogen Corporation,Carlsbad, Calif.) supplemented with 10% fetal calf serum (InvitrogenCorporation, Carlsbad, Calif.), penicillin 100 units per mL, andstreptomycin 100 micrograms per mL (Invitrogen Corporation, Carlsbad,Calif.). Cells were routinely passaged by trypsinization and dilutionwhen they reached 90% confluence.

[0200] NHDF Cells:

[0201] Human neonatal dermal fibroblast (NHDF) were obtained from theClonetics Corporation (Walkersville, Md.). NHDFs were routinelymaintained in Fibroblast Growth Medium (Clonetics Corporation,Walkersville, Md.) supplemented as recommended by the supplier. Cellswere maintained for up to 10 passages as recommended by the supplier.

[0202] HEK Cells:

[0203] Human embryonic keratinocytes (HEK) were obtained from theClonetics Corporation (Walkersville, Md.). HEKs were routinelymaintained in Keratinocyte Growth Medium (Clonetics Corporation,Walkersville, Md.) formulated as recommended by the supplier. Cells wereroutinely maintained for up to 10 passages as recommended by thesupplier.

[0204] b.END Cells:

[0205] The mouse brain endothelial cell line b.END was obtained from Dr.Werner Risau at the Max Plank Instititute (Bad Nauheim, Germany). b.ENDcells were routinely cultured in DMEM, high glucose (Gibco/LifeTechnologies, Gaithersburg, Md.) supplemented with 10% fetal calf serum(Gibco/Life Technologies, Gaithersburg, Md.). Cells were routinelypassaged by trypsinization and dilution when they reached 90%confluence. Cells were seeded into 96-well plates (Falcon-Primaria#3872) at a density of 3000 cells/well for use in RT-PCR analysis.

[0206] For Northern blotting or other analyses, cells may be seeded onto100 mm or other standard tissue culture plates and treated similarly,using appropriate volumes of medium and oligonucleotide.

[0207] Treatment with Antisense Compounds:

[0208] When cells reached 65-75% confluency, they were treated witholigonucleotide. For cells grown in 96-well plates, wells were washedonce with 100 μL OPTI-MEM™-1 reduced-serum medium (InvitrogenCorporation, Carlsbad, Calif.) and then treated with 130 μL ofOPTI-MEM™-1 containing 3.75 μg/mL LIPOFECTIN™ (Invitrogen Corporation,Carlsbad, Calif.) and the desired concentration of oligonucleotide.Cells are treated and data are obtained in triplicate. After 4-7 hoursof treatment at 37° C., the medium was replaced with fresh medium. Cellswere harvested 16-24 hours after oligonucleotide treatment.

[0209] The concentration of oligonucleotide used varies from cell lineto cell line. To determine the optimal oligonucleotide concentration fora particular cell line, the cells are treated with a positive controloligonucleotide at a range of concentrations. For human cells thepositive control oligonucleotide is selected from either ISIS 13920(TCCGTCATCGCTCCTCAGGG, SEQ ID NO: 1) which is targeted to human H-ras,or ISIS 18078, (GTGCGCGCGAGCCCGAAATC, SEQ ID NO: 2) which is targeted tohuman Jun-N-terminal kinase-2 (JNK2). Both controls are2′-O-methoxyethyl gapmers (2′-O-methoxyethyls shown in bold) with aphosphorothioate backbone. For mouse or rat cells the positive controloligonucleotide is ISIS 15770, ATGCATTCTGCCCCCAAGGA, SEQ ID NO: 3, a2′-O-methoxyethyl gapmer (2′-O-methoxyethyls shown in bold) with aphosphorothioate backbone which is targeted to both mouse and rat c-raf.The concentration of positive control oligonucleotide that results in80% inhibition of c-H-ras (for ISIS 13920), JNK2 (for ISIS 18078) orc-raf (for ISIS 15770) mRNA is then utilized as the screeningconcentration for new oligonucleotides in subsequent experiments forthat cell line. If 80% inhibition is not achieved, the lowestconcentration of positive control oligonucleotide that results in 60%inhibition of c-H-ras, JNK2 or c-raf mRNA is then utilized as theoligonucleotide screening concentration in subsequent experiments forthat cell line. If 60% inhibition is not achieved, that particular cellline is deemed as unsuitable for oligonucleotide transfectionexperiments. The concentrations of antisense oligonucleotides usedherein are from 50 nM to 300 nM.

Example 10 Analysis of Oligonucleotide Inhibition of HIF1α and/or HIF2αExpression

[0210] Antisense modulation of HIF1α and/or HIF2α expression can beassayed in a variety of ways known in the art. For example, HIF1α orHIF2α mRNA levels can be quantitated by, e.g., Northern blot analysis,competitive polymerase chain reaction (PCR), or real-time PCR (RT-PCR).Real-time quantitative PCR is presently preferred. RNA analysis can beperformed on total cellular RNA or poly(A)+ mRNA. The preferred methodof RNA analysis of the present invention is the use of total cellularRNA as described in other examples herein. Methods of RNA isolation arewell known in the art. Northern blot analysis is also routine in theart. Real-time quantitative (PCR) can be conveniently accomplished usingthe commercially available ABI PRISM™ 7600, 7700, or 7900 SequenceDetection System, available from PE-Applied Biosystems, Foster City,Calif. and used according to manufacturer's instructions.

[0211] Protein levels of HIF1α or HIF2α can be quantitated in a varietyof ways well known in the art, such as immunoprecipitation, Western blotanalysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA) orfluorescence-activated cell sorting (FACS). Antibodies directed to HIF1αor HIF2α can be identified and obtained from a variety of sources, suchas the MSRS catalog of antibodies (Aerie Corporation, Birmingham,Mich.), or can be prepared via conventional monoclonal or polyclonalantibody generation methods well known in the art.

Example 11 Design of Phenotypic Assays and In Vivo Studies for the Useof HIF1α or HIF2α Inhibitors

[0212] Phenotypic Assays

[0213] Once HIF1α or HIF2α inhibitors have been identified by themethods disclosed herein, the compounds are further investigated in oneor more phenotypic assays, each having measurable endpoints predictiveof efficacy in the treatment of a particular disease state or condition.

[0214] Phenotypic assays, kits and reagents for their use are well knownto those skilled in the art and are herein used to investigate the roleand/or association of HIF1α and/or HIF2α in health and disease.Representative phenotypic assays, which can be purchased from any one ofseveral commercial vendors, include those for determining cellviability, cytotoxicity, proliferation or cell survival (MolecularProbes, Eugene, Oreg.; PerkinElmer, Boston, Mass.), protein-based assaysincluding enzymatic assays (Panvera, LLC, Madison, Wis.; BD Biosciences,Franklin Lakes, N.J.; Oncogene Research Products, San Diego, Calif.),cell regulation, signal transduction, inflammation, oxidative processesand apoptosis (Assay Designs Inc., Ann Arbor, Mich.), triglycerideaccumulation (Sigma-Aldrich, St. Louis, Mo.), angiogenesis assays, tubeformation assays, cytokine and hormone assays and metabolic assays(Chemicon International Inc., Temecula, Calif.; Amersham Biosciences,Piscataway, N.J.).

[0215] In one non-limiting example, cells determined to be appropriatefor a particular phenotypic assay (i.e., MCF-7 cells selected for breastcancer studies; adipocytes for obesity studies) are treated with HIF1αand/or HIF2α inhibitors identified from the in vitro studies as well ascontrol compounds at optimal concentrations which are determined by themethods described above. At the end of the treatment period, treated anduntreated cells are analyzed by one or more methods specific for theassay to determine phenotypic outcomes and endpoints.

[0216] Phenotypic endpoints include changes in cell morphology over timeor treatment dose as well as changes in levels of cellular componentssuch as proteins, lipids, nucleic acids, hormones, saccharides ormetals. Measurements of cellular status which include pH, stage of thecell cycle, intake or excretion of biological indicators by the cell,are also endpoints of interest.

[0217] Analysis of the geneotype of the cell (measurement of theexpression of one or more of the genes of the cell) after treatment isalso used as an indicator of the efficacy or potency of the HIF1α and/orHIF2α inhibitors. Hallmark genes, or those genes suspected to beassociated with a specific disease state, condition, or phenotype, aremeasured in both treated and untreated cells.

Example 12 RNA Isolation

[0218] Poly(A)+ mRNA Isolation

[0219] Poly(A)+ mRNA was isolated according to Miura et al., (Clin.Chem., 1996, 42, 1758-1764). Other methods for poly(A)+ mRNA isolationare routine in the art. Briefly, for cells grown on 96-well plates,growth medium was removed from the cells and each well was washed with200 μL cold PBS. 60 μL lysis buffer (10 mM Tris-HCl, pH 7.6, 1 mM EDTA,0.5 M NaCl, 0.5% NP-40, 20 mM vanadyl-ribonucleoside complex) was addedto each well, the plate was gently agitated and then incubated at roomtemperature for five minutes. 55 μL of lysate was transferred to Oligod(T) coated 96-well plates (AGCT Inc., Irvine Calif.). Plates wereincubated for 60 minutes at room temperature, washed 3 times with 200 μLof wash buffer (10 mM Tris-HCl pH 7.6, 1 mM EDTA, 0.3 M NaCl). After thefinal wash, the plate was blotted on paper towels to remove excess washbuffer and then air-dried for 5 minutes, 60 μL of elution buffer (5 mMTris-HCl pH 7.6), preheated to 70° C., was added to each well, the platewas incubated on a 90° C. hot plate for 5 minutes, and the eluate wasthen transferred to a fresh 96-well plate.

[0220] Cells grown on 100 mm or other standard plates may be treatedsimilarly, using appropriate volumes of all solutions.

[0221] Total RNA Isolation

[0222] Total RNA was isolated using an RNEASY 96™ kit and bufferspurchased from Qiagen Inc. (Valencia, Calif.) following themanufacturer's recommended procedures. Briefly, for cells grown on96-well plates, growth medium was removed from the cells and each wellwas washed with 200 μL cold PBS. 150 μL Buffer RLT was added to eachwell and the plate vigorously agitated for 20 seconds. 150 μL of 70%ethanol was then added to each well and the contents mixed by pipettingthree times up and down. The samples were then transferred to the RNEASY96™ well plate attached to a QIAVAC™ manifold fitted with a wastecollection tray and attached to a vacuum source. Vacuum was applied for1 minute. 500 μL of Buffer RW1 was added to each well of the RNEASY 96™plate and incubated for 15 minutes and the vacuum was again applied for1 minute. An additional 500 μL of Buffer RW1 was added to each well ofthe RNEASY 96™ plate and the vacuum was applied for 2 minutes. 1 mL ofBuffer RPE was then added to each well of the RNEASY 96™ plate and thevacuum applied for a period of 90 seconds. The Buffer RPE wash was thenrepeated and the vacuum was applied for an additional 3 minutes. Theplate was then removed from the QIAVAC™ manifold and blotted dry onpaper towels. The plate was then re-attached to the QIAVAC™ manifoldfitted with a collection tube rack containing 1.2 mL collection tubes.RNA was then eluted by pipetting 140 μL of RNAse free water into eachwell, incubating 1 minute, and then applying the vacuum for 3 minutes.

[0223] The repetitive pipetting and elution steps may be automated usinga QIAGEN Bio-Robot 9604 (Qiagen, Inc., Valencia Calif.). Essentially,after lysing of the cells on the culture plate, the plate is transferredto the robot deck where the pipetting, DNase treatment and elution stepsare carried out.

Example 13 Real-Time Quantitative PCR Analysis of HIF1α mRNA Levels

[0224] Quantitation of HIF1α mRNA levels was accomplished by real-timequantitative PCR using the ABI PRISM™ 7600, 7700, or 7900 SequenceDetection System (PE-Applied Biosystems, Foster City, Calif.) accordingto manufacturer's instructions. This is a closed-tube, non-gel-based,fluorescence detection system which allows high-throughput quantitationof polymerase chain reaction (PCR) products in real-time. As opposed tostandard PCR in which amplification products are quantitated after thePCR is completed, products in real-time quantitative PCR are quantitatedas they accumulate. This is accomplished by including in the PCRreaction an oligonucleotide probe that anneals specifically between theforward and reverse PCR primers, and contains two fluorescent dyes. Areporter dye (e.g., FAM or JOE, obtained from either PE-AppliedBiosystems, Foster City, Calif., Operon Technologies Inc., Alameda,Calif. or Integrated DNA Technologies Inc., Coralville, Iowa) isattached to the 5′ end of the probe and a quencher dye (e.g., TAMRA,obtained from either PE-Applied Biosystems, Foster City, Calif., OperonTechnologies Inc., Alameda, Calif. or Integrated DNA Technologies Inc.,Coralville, Iowa) is attached to the 3′ end of the probe. When the probeand dyes are intact, reporter dye emission is quenched by the proximityof the 3′ quencher dye. During amplification, annealing of the probe tothe target sequence creates a substrate that can be cleaved by the5′-exonuclease activity of Taq polymerase. During the extension phase ofthe PCR amplification cycle, cleavage of the probe by Taq polymerasereleases the reporter dye from the remainder of the probe (and hencefrom the quencher moiety) and a sequence-specific fluorescent signal isgenerated. With each cycle, additional reporter dye molecules arecleaved from their respective probes, and the fluorescence intensity ismonitored at regular intervals by laser optics built into the ABI PRISM™Sequence Detection System. In each assay, a series of parallel reactionscontaining serial dilutions of mRNA from untreated control samplesgenerates a standard curve that is used to quantitate the percentinhibition after antisense oligonucleotide treatment of test samples.

[0225] Prior to quantitative PCR analysis, primer-probe sets specific tothe target gene being measured are evaluated for their ability to be“multiplexed” with a GAPDH amplification reaction. In multiplexing, boththe target gene and the internal standard gene GAPDH are amplifiedconcurrently in a single sample. In this analysis, mRNA isolated fromuntreated cells is serially diluted. Each dilution is amplified in thepresence of primer-probe sets specific for GAPDH only, target gene only(“single-plexing”), or both (multiplexing). Following PCR amplification,standard curves of GAPDH and target mRNA signal as a function ofdilution are generated from both the single-plexed and multiplexedsamples. If both the slope and correlation coefficient of the GAPDH andtarget signals generated from the multiplexed samples fall within 10% oftheir corresponding values generated from the single-plexed samples, theprimer-probe set specific for that target is deemed multiplexable. Othermethods of PCR are also known in the art.

[0226] PCR reagents were obtained from Invitrogen Corporation,(Carlsbad, Calif.). RT-PCR reactions were carried out by adding 20 μLPCR cocktail (2.5× PCR buffer minus MgCl₂, 6.6 mM MgCl₂, 375 μM each ofdATP, dCTP, dCTP and dGTP, 375 nM each of forward primer and reverseprimer, 125 nM of probe, 4 Units RNAse inhibitor, 1.25 Units PLATINUM®Taq, 5 Units MuLV reverse transcriptase, and 2.5× ROX dye) to 96-wellplates containing 30 μL total RNA solution (20-200 ng). The RT reactionwas carried out by incubation for 30 minutes at 48° C. Following a 10minute incubation at 95° C. to activate the PLATINUM® Taq, 40 cycles ofa two-step PCR protocol were carried out: 95° C. for 15 seconds(denaturation) followed by 60° C. for 1.5 minutes (annealing/extension).

[0227] Gene target quantities obtained by real time RT-PCR arenormalized using either the expression level of GAPDH, a gene whoseexpression is constant, or by quantifying total RNA using RiboGreen™(Molecular Probes, Inc. Eugene, Oreg.). GAPDH expression is quantifiedby real time RT-PCR, by being run simultaneously with the target,multiplexing, or separately. Total RNA is quantified using RiboGreen™RNA quantification reagent (Molecular Probes, Inc. Eugene, Oreg.).Methods of RNA quantification by RiboGreen™ are taught in Jones, L. J.,et al, (Analytical Biochemistry, 1998, 265, 368-374).

[0228] In this assay, 170 μL of RiboGreen™ working reagent (RiboGreen™reagent diluted 1:350 in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5) is pipettedinto a 96-well plate containing 30 μL purified, cellular RNA. The plateis read in a CytoFluor 4000 (PE Applied Biosystems) with excitation at485 nm and emission at 530 nm.

[0229] Probes and primers to human HIF1α were designed to hybridize to ahuman HIF1α sequence, using published sequence information (GenBankaccession number U29165.1, incorporated herein by reference andincorporated herein as SEQ ID NO:4). For human HIF1α the PCR primerswere:

[0230] forward primer: CCAGTTACGTTCCTTCGATCAGT (SEQ ID NO: 5) reverseprimer: TTTGAGGACTTGCGCTTTCA (SEQ ID NO: 6) and the PCR probe was:FAM-TCACCATTAGAAAGCAGTTCCGCAAGCC-TAMRA (SEQ ID NO: 7) where FAM is thefluorescent dye and TAMRA is the quencher dye. For human GAPDH the PCRprimers were:

[0231] forward primer: GAAGGTGAAGGTCGGAGTC(SEQ ID NO:8) reverse primer:GAAGATGGTGATGGGATTTC (SEQ ID NO:9) and the PCR probe was: 5′JOE-CAAGCTTCCCGTTCTCAGCC-TAMRA 3′ (SEQ ID NO: 10) where JOE is thefluorescent reporter dye and TAMRA is the quencher dye.

Example 14 Northern Blot Analysis of HIF1α mRNA Levels

[0232] Eighteen hours after antisense treatment, cell monolayers werewashed twice with cold PBS and lysed in 1 mL RNAZOL™ (TEL-TEST “B” Inc.,Friendswood, Tex.). Total RNA was prepared following manufacturer'srecommended protocols. Twenty micrograms of total RNA was fractionatedby electrophoresis through 1.2% agarose gels containing 1.1%formaldehyde using a MOPS buffer system (AMRESCO, Inc. Solon, Ohio). RNAwas transferred from the gel to HYBOND™-N+ nylon membranes (AmershamPharmacia Biotech, Piscataway, N.J.) by overnight capillary transferusing a Northern/Southern Transfer buffer system (TEL-TEST “B” Inc.,Friendswood, Tex.). RNA transfer was confirmed by UV visualization.Membranes were fixed by UV cross-linking using a STRATALINKER™ UVCrosslinker 2400 (Stratagene, Inc, La Jolla, Calif.) and then probedusing QUICKHYB™ hybridization solution (Stratagene, La Jolla, Calif.)using manufacturer's recommendations for stringent conditions.

[0233] To detect human HIF1α, a human HIF1α specific probe was preparedby PCR using the forward primer CCAGTTACGTTCCTTCGATCAGT (SEQ ID NO: 5)and the reverse primer TTTGAGGACTTGCGCTTTCA (SEQ ID NO: 6). To normalizefor variations in loading and transfer efficiency membranes werestripped and probed for human glyceraldehyde-3-phosphate dehydrogenase(GAPDH) RNA (Clontech, Palo Alto, Calif.).

[0234] Hybridized membranes were visualized and quantitated using aPHOSPHORIMAGER™ and IMAGEQUANT™ Software V3.3 (Molecular Dynamics,Sunnyvale, Calif.). Data was normalized to GAPDH levels in untreatedcontrols.

Example 15 Antisense Inhibition of Human HIF1α Expression by ChimericPhosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap

[0235] In accordance with the present invention, a series of antisensecompounds were designed to target different regions of the human HIF1αRNA, using published sequences (GenBank accession number U29165.1,incorporated herein by reference and incorporated herein as SEQ ID NO:4, positions 82000 to 139500 of the sequence with GenBank accessionnumber AL137129.4, incorporated herein by reference and incorporatedherein as SEQ ID NO: 11, GenBank accession number AU123241.1,incorporated herein by reference and incorporated herein as SEQ ID NO:12, and GenBank accession number AB073325.1, incorporated herein byreference and incorporated herein as SEQ ID NO: 13). The compounds areshown in Table 1. “Target site” indicates the first (5′-most) nucleotidenumber on the particular target sequence to which the compound binds.All compounds in Table 1 are chimeric oligonucleotides (“gapmers”) 20nucleotides in length, composed of a central “gap” region consisting often 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′directions) by five-nucleotide “wings”. The wings are composed of2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone)linkages are phosphorothioate (P═S) throughout the oligonucleotide. Allcytidine residues are 5-methylcytidines. The compounds were analyzed fortheir effect on human HIF1α mRNA levels by quantitative real-time PCR asdescribed in other examples herein. Data are averages from threeexperiments in which A549 cells were treated with the antisenseoligonucleotides of the present invention. If present, “N.D.” indicates“no data”. TABLE 1 Inhibition of human HIF1α mRNA levels by chimericphosphorothioate oligonucleotides having 2′-MOE wings and a deoxy gapTARGET SEQ ID TARGET % SEQ ID ISIS # REGION NO SITE SEQUENCE INHIB NO175477 Coding 4 2496 aaagtgatgtagtagctgca 54 14 175478 Coding 4 854ggtatcatatacgtgaatgt 73 15 175479 3′UTR 4 3179 taccacgtactgctggcaaa 3116 175480 Coding 4 2039 tgtgctttgaggacttgcgc 94 17 175481 Coding 4 583gaaatgtaaatcatgtcacc 56 18 175482 Coding 4 1408 tcaaagaggctacttgtatc 7519 175483 Coding 4 1674 ttaatgcaacttcttgattg 45 20 175484 3′UTR 4 3333atcattattatatgattaac 60 21 175485 5′UTR 4 152 gaaaggcaagtccagaggtg 42 22175486 3′UTR 4 3027 taaactccctagccaaaaat 40 23 175487 Coding 4 2085cattagcagtaggttcttgt 75 24 175488 3′UTR 4 3101 gatcatgatgaaaggttact 8625 175489 Coding 4 1001 aaatttcatatccaggctgt 85 26 175490 Coding 4 460agtttcctcacacgcaaata 38 27 175491 Coding 4 1983 actgatcgaaggaacgtaac 8728 175492 Coding 4 2404 cgctttctctgagcattctg 44 29 175493 Coding 4 649aaatcaaacacactgtgtcc 79 30 175494 Coding 4 1139 tcctttagtaaacatatcat 7131 175495 Coding 4 1442 caaagttaaagcatcaggtt 79 32 175496 Coding 4 1765ctagtgcttccatcggaagg 37 33 175497 3′UTR 4 3424 aatgccacataccttctaga 2434 175498 5′UTR 4 110 tcgtgagactagagagaagc 71 35 175499 3′UTR 4 3094atgaaaggttactgccttct 81 36 175500 Coding 4 912 tcagcaccaagcaggtcata 8 37175501 3′UTR 4 2841 aagtttgtgcagtattgtag 33 38 175502 Coding 4 2396ctgagcattctgcaaagcta 0 39 175503 Coding 4 350 ttcagattctttacttcgcc 54 40175504 Coding 4 2320 gataacacgttagggcttct 41 41 175505 Coding 4 2331tcaaagcgacagataacacg 51 42 175506 Coding 4 1091 caaagcatgataatattcat 5643 175507 Coding 4 565 ccatcatctgtgagaaccat 86 44 175508 Coding 4 2222atatggtgatgatgtggcac 76 45 175509 5′UTR 4 51 ctcctcaggtggcttgtcag 33 46175510 3′UTR 4 2931 tgagctgtctgtgatccagc 94 47 175511 Coding 4 2321agataacacgttagggcttc 86 48 175512 Start 4 248 catggtgaatcggtccccgc 76 49Codon 175513 Coding 4 1224 tgttatatatgacagttgct 73 50 224184 Coding 4414 ccttatcaagatgcgaactc 63 51 224185 Coding 4 480 ccaaatcaccagcatccaga32 52 224186 Coding 4 619 aactgagttaatcccatgta 72 53 224187 Coding 4 627ttagttcaaactgagttaat 31 54 224188 Coding 4 706 aggccatttctgtgtgtaag 6255 224189 Coding 4 961 ctatctaaaggaatttcaat 10 56 224190 Coding 4 1036cccatcaattcggtaattct 41 57 224191 Coding 4 1125 tatcatgatgagttttggtc 8158 224192 Coding 4 1283 aataataccactcacaacgt 60 59 224193 Coding 4 1380caactttggtgaatagctga 71 60 224194 Coding 4 1699 agtgactctggatttggttc 4461 224195 Coding 4 1928 catctccaagtctaaatctg 36 62 224196 Coding 4 1995ctaatggtgacaactgatcg 72 63 224197 Coding 4 2126 cactgtttttaattcatcag 6564 224198 Coding 4 2457 ataatgttccaattcctact 31 65 224199 Stop 4 2735agaaaaagctcagttaactt 57 66 Codon 224200 3′UTR 4 2828attgtagccaggcttctaaa 68 67 224201 3′UTR 4 3056 atcttcttaaaaataattcg 1868 224202 3′UTR 4 3193 tgtgcaattgtggctaccac 76 69 224203 3′UTR 4 3316aacaatgtcatgttccaggt 88 70 224204 3′UTR 4 3486 gctggcaaagtgactataga 7271 224205 3′UTR 4 3896 ttccacagaagatgtttatt 30 72 224206 3′UTR 4 3899tttttccacagaagatgttt 14 73 224207 intron 11 11258 tagagctaaacgatctagaa47 74 224208 intron 11 23630 taactctttctggccttgaa 93 75 224209 intron 1125682 attggccctaacagaaaatc 19 76 224210 intron: 11 27616agaacttatcctacttaaca 7 77 exon junction 224211 intron 11 39357gtttccctcgtgttgctcag 63 78 224212 exon: 11 39759 ttgtacttactatcatgatg 2579 intron junction 224213 exon: 11 41520 acttacttacctcacaacgt 9 80intron junction 224214 intron: 11 47989 aatctgtgtcctttaaaaca 35 81 exonjunction 224215 exon 11 2745 tgtgcactgaggagctgagg 19 82 224216 exon 4296 acgttcagaacttatctttt 45 83 224217 Stop 13 2221 catgctaaataattcctact0 84 Codon

[0236] As shown in Table 1, SEQ ID NOs 14, 15, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 28, 29, 30, 31, 32, 35, 36, 40, 41, 42, 43, 44, 45, 47,48, 49, 50, 51, 53, 55, 57, 58, 59, 60, 61, 63, 64, 66, 67, 69, 70, 71,74, 75, 78 and 83 demonstrated at least 40% inhibition of human HIF1αexpression in this assay and are therefore preferred. More preferred areSEQ ID NOs 47, 48 and 25. The target regions to which these preferredsequences are complementary are herein referred to as “preferred targetsegments” and are therefore preferred for targeting by compounds of thepresent invention. These preferred target segments are shown in Table 2.The sequences represent the reverse complement of the preferredantisense compounds shown in Table 1. “Target site” indicates the first(5′-most) nucleotide number on the particular target nucleic acid towhich the oligonucleotide binds. Also shown in Table 2 is the species inwhich each of the preferred target segments was found. TABLE 2 Sequenceand position of preferred target segments identified in HIF1α. TARGETSITE SEQ ID TARGET REV COMP SEQ ID ID NO SITE SEQUENCE OF SEQ ID ACTIVEIN NO 90592 4 2496 tgcagctactacatcacttt 14 H. sapiens 85 90593 4 854acattcacgtatatgatacc 15 H. sapiens 86 90595 4 2039 gcgcaagtcctcaaagcaca17 H. sapiens 87 90596 4 583 ggtgacatgatttacatttc 18 H. sapiens 88 905974 1408 gatacaagtagcctctttga 19 H. sapiens 89 90598 4 1674caatcaagaagttgcattaa 20 H. sapiens 90 90599 4 3333 gttaatcatataataatgat21 H. sapiens 91 90600 4 152 cacctctggacttgcctttc 22 H. sapiens 92 906014 3027 atttttggctagggagttta 23 H. sapiens 93 90602 4 2085acaagaacctactgctaatg 24 H. sapiens 94 90603 4 3101 agtaacctttcatcatgatc25 H. sapiens 95 90604 4 1001 acagcctggatatgaaattt 26 H. sapiens 9690606 4 1983 gttacgttccttcgatcagt 28 H. sapiens 97 90607 4 2404cagaatgctcagagaaagcg 29 H. sapiens 98 90608 4 649 ggacacagtgtgtttgattt30 H. sapiens 99 90609 4 1139 atgatatgtttactaaagga 31 H. sapiens 10090610 4 1442 aacctgatgctttaactttg 32 H. sapiens 101 90613 4 110gcttctctctagtctcacga 35 H. sapiens 102 90614 4 3094 agaaggcagtaacctttcat36 H. sapiens 103 90618 4 350 ggcgaagtaaagaatctgaa 40 H. sapiens 10490619 4 2320 agaagccctaacgtgttatc 41 H. sapiens 105 90620 4 2331cgtgttatctgtcgctttga 42 H. sapiens 106 90621 4 1091 atgaatattatcatgctttg43 H. sapiens 107 90622 4 565 atggttctcacagatgatgg 44 H. sapiens 10890623 4 2222 gtgccacatcatcaccatat 45 H. sapiens 109 90625 4 2931gctggatcacagacagctca 47 H. sapiens 110 90626 4 2321 gaagccctaacgtgttatct48 H. sapiens 111 90627 4 248 gcggggaccgattcaccatg 49 H. sapiens 11290628 4 1224 agcaactgtcatatataaca 50 H. sapiens 113 140838 4 414gagttcgcatcttgataagg 51 H. sapiens 114 140840 4 619 tacatgggattaactcagtt53 H. sapiens 115 140842 4 706 cttacacacagaaatggcct 55 H. sapiens 116140844 4 1036 agaattaccgaattgatggg 57 H. sapiens 117 140845 4 1125gaccaaaactcatcatgata 58 H. sapiens 118 140846 4 1283acgttgtgagtggtattatt 59 H. sapiens 119 140847 4 1380tcagctattcaccaaagttg 60 H. sapiens 120 140848 4 1699gaaccaaatccagagtcact 61 H. sapiens 121 140850 4 1995cgatcagttgtcaccattag 63 H. sapiens 122 140851 4 2126ctgatgaattaaaaacagtg 64 H. sapiens 123 140853 4 2735aagttaactgagctttttct 66 H. sapiens 124 140854 4 2828tttagaagcctggctacaat 67 H. sapiens 125 140856 4 3193gtggtagccacaattgcaca 69 H. sapiens 126 140857 4 3316acctggaacatgacattgtt 70 H. sapiens 127 140858 4 3486tctatagtcactttgccagc 71 H. sapiens 128 140861 11 11258ttctagatcgtttagctcta 74 H. sapiens 129 140862 11 23630ttcaaggccagaaagagtta 75 H. sapiens 130 140865 11 39357ctgagcaacacgagggaaac 78 H. sapiens 131 140870 4 296 aaaagataagttctgaacgt83 H. sapiens 132

[0237] As these “preferred target segments” have been found byexperimentation to be open to, and accessible for, hybridization withthe antisense compounds of the present invention, one of skill in theart will recognize or be able to ascertain, using no more than routineexperimentation, further embodiments of the invention that encompassother compounds that specifically hybridize to these preferred targetsegments and consequently inhibit the expression of HIF1α.

[0238] According to the present invention, antisense compounds includeantisense oligomeric compounds, antisense oligonucleotides, ribozymes,external guide sequence (EGS) oligonucleotides, alternate splicers,primers, probes, and other short oligomeric compounds which hybridize toat least a portion of the target nucleic acid.

Example 16 Western Blot Analysis of HIF1α or HIF2α Prot in Levels

[0239] Western blot analysis (immunoblot analysis) is carried out usingstandard methods. Cells are harvested 16-20 h after oligonucleotidetreatment, washed once with PBS, suspended in Laemmli buffer (100ul/well), boiled for 5 minutes and loaded on a 16% SDS-PAGE gel. Gelsare run for 1.5 hours at 150 V, and transferred to membrane for westernblotting. Appropriate primary antibody directed to HIF1α or HIF2α isused, with a radiolabeled or fluorescently labeled secondary antibodydirected against the primary antibody species. Bands are visualizedusing a PHOSPHORIMAGER™ (Molecular Dynamics, Sunnyvale Calif.).

Example 17 Additional Antisense Oligonucleotides Against Human HIF1α

[0240] A series of antisense compounds were designed to target differentregions of the human HIF1α RNA, using published sequences (GenBankaccession number U29165.1, incorporated herein by reference andincorporated herein as SEQ ID NO: 133). The compounds are shown in Table3. “Target site” indicates the first (5′-most) nucleotide number on theparticular target sequence to which the compound binds. All compounds inTable 3 are chimeric oligonucleotides (“gapmers”) 20 nucleotides inlength, composed of a central “gap” region consisting of ten2′-deoxynucleotides, which is flanked on both sides (5′ and 3′directions) by five-nucleotide “wings”. The wings are composed of2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone)linkages are phosphorothioate (P═S) throughout the oligonucleotide. Allcytidine residues are 5-methylcytidines. The compounds were analyzed fortheir effect on human HIF1α mRNA levels by quantitative real-time PCR asdescribed in other examples herein. Data are averages from threeexperiments in which A549 cells were treated with the antisenseoligonucleotides of the present invention. “Species” indicates theanimal species of HIF1α nucleic acid to which the compounds are fullycomplementary (H=human, M=mouse, R=rat). As noted many of the compoundsare fully complementary to more than one species. TABLE 3 Inhibition ofhuman HIF1α MrNA levels by additional chimeric phosphorothioateoligonucleotides having 2′-MOE wings and a deoxy gap TARGET seq idTARGET SEQ ISIS # REGION no site Sequence % INHIB ID NO Species 298690Coding 133 373 tgatgagcaagctcataaaa 51 134 H, M, R 298692 Coding 133 378gcaactgatgagcaagctca 77 135 H, M, R 298692 Coding 133 385ggaagtggcaactgatgagc 62 136 H, M, R 298693 Coding 133 631ccagttagttcaaactgagt 79 137 H, M, R 298694 Coding 133 636tgtgtccagttagttcaaac 79 138 H, M, R 298695 Coding 133 641cacactgtgtccagttagtt 79 139 H, M, R 298696 Coding 133 663cacatggatgagtaaaatca 69 140 H, M 298697 Coding 133 673tcctcatggtcacatggatg 84 141 H, M, R 298698 Coding 133 682tctctcatttcctcatggtc 80 142 H, M, R 298699 Coding 133 687gcatttctctcatttcctca 73 143 H, M, R 298700 Coding 133 695gtgtgtaagcatttctctca 67 144 H, M, R 298701 Coding 133 705ggccatttctgtgtgtaagc 78 145 H, M, R 298702 Coding 133 865tggttactgttggtatcata 85 146 H, M 298703 Coding 133 919tcacaaatcagcaccaagca 57 147 H, M, R 298704 Coding 133 924tgggttcacaaatcagcacc 71 148 H, M, R 298705 Coding 133 931tgaggaatgggttcacaaat 69 149 H, M, R 298706 Coding 133 967gtcttgctatctaaaggaat 58 150 H, M 298707 Coding 133 1078tattcataaattgagcggcc 80 151 H, M 298708 Coding 133 1084tgataatattcataaattga 13 152 H, M, R 298709 Coding 133 1117tgagttttggtcagatgatc 64 153 H, M, R 298710 Coding 133 1144acttgtcctttagtaaacat 58 154 H, M, R 298711 Coding 133 1149tggtgacttgtcctttagta 75 155 H, M,R 298712 Coding 133 1154tcctgtggtgacttgtcctt 76 156 H, M, R 298713 Coding 133 1159tactgtcctgtggtgacttg 62 157 H, M, R 298714 Coding 133 1164tcctgtactgtcctgtggtg 83 158 H, M, R 298715 Coding 133 1171gcaagcatcctgtactgtcc 67 159 H, M, R 298716 Coding 133 1192cagacatatccacctctttt 56 160 H, M, R 298717 Coding 133 1198tcaacccagacatatccacc 53 161 H, M, R 298718 Coding 133 1217tatgacagttgcttgagttt 64 162 H, M 298719 Coding 133 1222ttatatatgacagttgcttg 69 163 H, M 298720 Coding 133 1308gaagggagaaaatcaagtcg 46 164 H, M, R 298721 Coding 133 1320attctgtttgttgaagggag 43 165 H, M, R 298722 Coding 133 1354ttcatatctgaagattcaac 53 166 H, M, R 298723 Coding 133 1387tctgattcaactttggtgaa 59 167 H, M 298724 Coding 133 1549attacatcattatataatgg 39 168 H, M 298725 Coding 133 1639ctacttcgaagtggctttgg 77 169 H, M, R 298726 Coding 133 1645tcagcactacttcgaagtgg 80 170 H, M, R 298727 Coding 133 1771ctttgtctagtgcttccatc 73 171 H, M, R 298728 Coding 133 1955atcatccattgggatatagg 74 172 H, M, R 298729 Coding 133 1996tctaatggtgacaactgatc 78 173 H, M, R 298730 Coding 133 2421catcatgttccatttttcgc 69 174 H, M, R 298731 Coding 133 2632gtcagctgtggtaatccact 69 175 H, M, R 298732 Coding 133 2638taactggtcagctgtggtaa 58 176 H, M, R 298733 Coding 133 2659ggagcattaacttcacaatc 39 177 H, M, R 298734 Coding 133 2680aggtttctgctgccttgtat 65 178 H, M, R 298735 Coding 133 2689ccctgcagtaggtttctgct 63 179 H, M, R 298736 Coding 133 2694cttcaccctgcagtaggttt 76 180 H, M, R 298737 Coding 133 2699taattcttcaccctgcagta 71 181 H, M, R 298738 Coding 133 2704ctgagtaattcttcaccctg 77 182 H, M, R 298739 Coding 133 2709aagctctgagtaattcttca 84 183 H, M, R 298740 Coding 133 2714atccaaagctctgagtaatt 66 184 H, M, R 298741 Coding 133 2719acttgatccaaagctctgag 72 185 H, M, R 298742 Stop 133 2728gctcagttaacttgatccaa 80 186 H, M, R codon 298743 3′UTR 133 2770tgagccaccagtgtccaaaa 85 187 H, M, R 298744 3′UTR 133 2821ccaggcttctaaaattagat 68 188 H, M 298745 3′UTR 133 2835gtgcagtattgtagccaggc 78 189 H, M 298746 3′UTR 133 2840agtttgtgcagtattgtagc 74 190 H, M 298747 3′UTR 133 3004taaataaaaaggtgcatttt 0 191 H, M, R 298749 3′UTR 133 3110actgcctatgatcatgatga 74 192 H, M 298750 3′UTR 133 3194ttgtgcaattgtggctacca 79 193 H, M, R 298751 3′UTR 133 3199atatattgtgcaattgtggc 0 194 H, M, R 298752 3′UTR 133 3204agaaaatatattgtgcaatt 31 195 H, M, R 298753 3′UTR 133 3264cttaaaaactagttttataa 21 196 H, M, R 298754 3′UTR 133 3382atgtaaatggctttacccat 68 197 H, M, R 298755 3′UTR 133 3437ttttatccaaataaatgcca 59 198 H, M, R 298756 3′UTR 133 3443tgagaattttatccaaataa 44 199 H, M, R 298757 3′UTR 133 3701taatagcgacaaagtgcata 81 200 H, M, R 298758 3′UTR 133 3706gatgttaatagcgacaaagt 54 201 H, M, R 298759 3′UTR 133 3711aaaaggatgttaatagcgac 77 202 H, M, R 298760 3′UTR 133 3752aatgcttctaaaattactca 62 203 H, M, R 298761 3′UTR 133 3766tatattcctaaaataatgct 30 204 H, M 298762 3′UTR 133 3892acagaagatgtttatttgat 44 205 H, M, R

[0241] In Table 3, SEQ ID NO 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 166, 167, 169, 170, 171, 172, 173,174, 175, 176, 178, 179, 180, 181, 182, 184, 185, 186, 187, 188, 189,190, 192, 193, 197, 198, 200, 201, 202 and 203 demonstrated at least 50%inhibition of HIF1α expression and are therefore preferred.

Example 18 Antisense Inhibition of Mouse HIF1α Expression by ChimericPhosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap

[0242] In accordance with the present invention, a series of antisensecompounds were designed to target different regions of the mouse HIF1αRNA, using published sequences (GenBank accession number NM_(—)010431.1,incorporated herein by reference and incorporated herein as SEQ ID NO:206. The compounds are shown in Table 4. “Target site” indicates thefirst (5′-most) nucleotide number on the particular target sequence towhich the compound binds. All compounds in Table 4 are chimericoligonucleotides (“gapmers”) 20 nucleotides in length, composed of acentral “gap” region consisting of ten 2′-deoxynucleotides, which isflanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”.The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. Theinternucleoside (backbone) linkages are phosphorothioate (P═S)throughout the oligonucleotide. All cytidine residues are5-methylcytidines. The compounds were analyzed for their effect on mouseHIF1α mRNA levels by quantitative real-time PCR as described in otherexamples herein. Unlike previous examples, the oligonucleotideconcentration in this experiment is 50 nM. Data are averages from threeexperiments in which b.END cells were treated with the antisenseoligonucleotides of the present invention. In Table 4, “Species”indicates the animal species of HIF1α nucleic acid to which thecompounds are fully complementary (H=human, M=mouse, R=rat). As notedmany of the compounds are fully complementary to more than one species.TABLE 4 Inhibition of mouse HIF1α mRNA levels by chimericphosphorothioate oligonucleotides having 2′-MOE wings and a deoxy gapTARGET TARGET SEQ ISIS # REGION SEQ ID SITE Sequence % INHIB ID NOSpecies 298690 Coding 206 366 tgatgagcaagctcataaaa 32 134 H, M, R 298691Coding 206 371 gcaactgatgagcaagctca 67 135 H, M, R 298692 Coding 206 378ggaagtggcaactgatgagc 33 136 H, M, R 298693 Coding 206 624ccagttagttcaaactgagt 58 137 H, M, R 298694 Coding 206 629tgtgtccagttagttcaaac 39 138 H, M, R 298695 Coding 206 634cacactgtgtccagttagtt 71 139 H, M, R 298696 Coding 206 656cacatggatgagtaaaatca 60 140 H, M 298697 Coding 206 666tcctcatggtcacatggatg 56 141 H, M, R 298698 Coding 206 675tctctcatttcctcatggtc 69 142 H, M, R 298699 Coding 206 680gcatttctctcatttcctca 70 143 H, M, R 298700 Coding 206 688gtgtgtaagcatttctctca 64 144 H, M, R 298701 Coding 206 698ggccatttctgtgtgtaagc 46 145 H, M, R 298702 Coding 206 858tggttactgttggtatcata 69 146 H, M 298703 Coding 206 912tcacaaatcagcaccaagca 45 147 H, M, R 298704 Coding 206 917tgggttcacaaatcagcacc 34 148 H, M, R 298705 Coding 206 924tgaggaatgggttcacaaat 64 149 H, M, R 298706 Coding 206 960gtcttgctatctaaaggaat 42 150 H, M 298707 Coding 206 1071tattcataaattgagcggcc 64 151 H, M 298708 Coding 206 1077tgataatattcataaattga 0 152 H, M, R 298709 Coding 206 1110tgagttttggtcagatgatc 26 153 H, M, R 298710 Coding 206 1137acttgtcctttagtaaacat 47 154 H, M, R 298711 Coding 206 1142tggtgacttgtcctttagta 64 155 H, M, R 298712 Coding 206 1147tcctgtggtgacttgtcctt 58 156 H, M, R 298713 Coding 206 1152tactgtcctgtggtgacttg 48 157 H, M, R 298714 Coding 206 1157tcctgtactgtcctgtggtg 61 158 H, M, R 298715 Coding 206 1164gcaagcatcctgtactgtcc 70 159 H, M, R 298716 Coding 206 1185cagacatatccacctctttt 43 160 H, M, R 298717 Coding 206 1191tcaacccagacatatccacc 55 161 H, M, R 298718 Coding 206 1210tatgacagttgcttgagttt 39 162 H, M 298719 Coding 206 1215ttatatatgacagttgcttg 42 163 H, M 298720 Coding 206 1301gaagggagaaaatcaagtcg 23 164 H, M, R 298721 Coding 206 1313attctgtttgttgaagggag 30 165 H, M, R 298722 Coding 206 1347ttcatatctgaagattcaac 5 166 H, M, R 298723 Coding 206 1380tctgattcaactttggtgaa 52 167 H, M 298724 Coding 206 1542attacatcattatataatgg 29 168 H, M 298725 Coding 206 1629ctacttcgaagtggctttgg 57 169 H, M, R 298726 Coding 206 1635tcagcactacttcgaagtgg 59 170 H, M, R 298727 Coding 206 1761ctttgtctagtgcttccatc 46 171 H, M, R 298728 Coding 206 1987atcatccattgggatatagg 29 172 H, M, R 298729 Coding 206 2028tccaatggtgacaactgatc 19 173 H, M, R 298730 Coding 206 2444catcatgttccatttttcgc 55 174 H, M, R 298731 Coding 206 2655gtcagctgtggtaatccact 59 175 H, M, R 298732 Coding 206 2661taactggtcagctgtggtaa 62 176 H, M, R 298733 Coding 206 2682ggagcattaacttcacaatc 32 177 H, M, R 298734 Coding 206 2703aggtttctgctgccttgtat 50 178 H, M, R 298735 Coding 206 2712ccctgcagtaggtttctgct 53 179 H, M, R 298736 Coding 206 2717cttcaccctgcagtaggttt 46 180 H, M, R 298737 Coding 206 2722taattcttcaccctgcagta 42 181 H, M, R 298738 Coding 206 2727ctgagtaattcttcaccctg 62 182 H, M, R 298739 Coding 206 2732aagctctgagtaattcttca 44 183 H, M, R 298740 Coding 206 2737atccaaagctctgagtaatt 42 184 H, M, R 298741 Coding 206 2742acttgatccaaagctctgag 47 185 H, M, R 298742 Stop 206 2751gctcagttaacttgatccaa 67 186 H, M, R codon 298743 3′UTR 206 2853tgagccaccagtgtccaaaa 56 187 H, M, R 298744 3′UTR 206 2895ccaggcttctaaaattagat 48 188 H, M 298745 3′UTR 206 2909gtgcagtattgtagccaggc 72 189 H, M 298746 3′UTR 206 2914agtttgtgcagtattgtagc 62 190 H, M 298747 3′UTR 206 3067taaataaaaaggtgcatttt 4 191 H, M, R 298748 3′UTR 206 3162gatcatgatgagaatttact 56 207 M 298749 3′UTR 206 3171 actgcctatgatcatgatga64 192 H, M, 298750 3′UTR 206 3253 ttgtgcaattgtggctacca 74 193 H, M, R298751 3′UTR 206 3258 atatattgtgcaattgtggc 67 194 H, M, R 298752 3′UTR206 3263 agaaaatatattgtgcaatt 24 195 H, M, R 298753 3′UTR 206 3322cttaaaaactagttttataa 0 196 H, M, R 298754 3′UTR 206 3428atgtaaatggctttacccat 51 197 H, M, R 298755 3′UTR 206 3483ttttatccaaataaatgcca 28 198 H, M, R 298756 3′UTR 206 3489tgagaattttatccaaataa 14 199 H, M, R 298757 3′UTR 206 3739taatagcgacaaagtgcata 43 200 H, M, R 298758 3′UTR 206 3744gatgttaatagcgacaaagt 23 201 H, M, R 298759 3′UTR 206 3749aaaaggatgttaatagcgac 45 202 H, M, R 298760 3′UTR 206 3789aatgcttctaaaattactca 30 203 H, M, R 298761 3′UTR 206 3803tatattcctaaaataatgct 0 204 H, M 298762 3′UTR 206 3928acagaagatgtttatttgat 21 205 H, M, R

[0243] In Table 4, SEQ ID NOs 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 154, 155, 156, 157,158, 159, 160, 161, 162, 163, 167, 169, 170, 171, 174, 175, 176, 177,178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 207,192, 193, 194, 197, 200, and 202 demonstrated at least 32% inhibition ofHIF1α expression and are therefore preferred.

Example 19 Real-Time Quantitative PCR Analysis of HIF2α mRNA Levels

[0244] Quantitation of HIF2α mRNA levels was accomplished by real-timequantitative PCR using the ABI PRISM™ 7600, 7700, or 7900 SequenceDetection System (PE-Applied Biosystems, Foster City, Calif.) asdescribed in previous examples.

[0245] Probes and primers to human HIF2α were designed to hybridize to ahuman HIF2α sequence, using published sequence information (GenBankaccession number NM_(—)001430.1, incorporated herein by reference andincorporated herein as SEQ ID NO: 208). For human HIF2α the PCR primerswere:

[0246] forward primer: AAGCCTTGGAGGGTTTCATTG (SEQ ID NO: 209) reverseprimer: TGCTGATGTTTTCTGACAGAAAGAT (SEQ ID NO: 210) and the PCR probewas: FAM-CGTGGTGACCCAAGATGGCGACA-TAMRA (SEQ ID NO: 211) where FAM is thefluorescent dye and TAMRA is the quencher dye. For human GAPDH the PCRprimers and probe were those listed in previous examples (SEQ ID NOs: 8,9, 10).

[0247] Probes and primers to mouse HIF2α were designed to hybridize to amouse HIF2α sequence, using published sequence information (GenBankaccession number NM_(—)010137.1, incorporated herein by reference andincorporated herein as SEQ ID NO: 212). For mouse HIF2α the PCR primerswere:

[0248] forward primer: GGCCATCGTTCGAGCCTTA (SEQ ID NO: 213) reverseprimer: GGCACGGGCACGTTCA (SEQ ID NO: 214) and the PCR probe was:FAM-CTGTTGCCGGAACTGACCAGATATGACTG-TAMRA (SEQ ID NO: 215) where FAM isthe fluorescent reporter dye and TAMRA is the quencher dye. For mouseGAPDH the PCR primers were:

[0249] forward primer: GGCAAATTCAACGGCACAGT(SEQ ID NO: 216) reverseprimer: GGGTCTCGCTCCTGGAAGAT(SEQ ID NO: 217) and the PCR probe was:5′JOE-AGGCCGAGAATGGGAAGCTTGTCATC-TAMRA 3′ (SEQ ID NO: 218) where JOE isthe fluorescent reporter dye and TAMRA is the quencher dye.

Example 20 Northern Blot Analysis of HIF2α mRNA Levels

[0250] Eighteen hours after antisense treatment, cell monolayers werewashed twice with cold PBS and lysed in 1 mL RNAZOL™ (TEL-TEST “B” Inc.,Friendswood, Tex.). Total RNA was prepared following manufacturer'srecommended protocols. Electrophoresis and blotting was performed asdescribed in previous examples.

[0251] To detect human HIF2α, a human HIF2α specific probe was preparedby PCR using the forward primer AAGCCTTGGAGGGTTTCATTG (SEQ ID NO: 209)and the reverse primer TGCTGATGTTTTCTGACAGAAAGAT (SEQ ID NO: 210). Tonormalize for variations in loading and transfer efficiency membraneswere stripped and probed for human glyceraldehyde-3-phosphatedehydrogenase (GAPDH) RNA (Clontech, Palo Alto, Calif.).

[0252] To detect mouse HIF2α, a mouse HIF2α specific probe was preparedby PCR using the forward primer GGCCATCGTTCGAGCCTTA (ID NO: 213) and thereverse primer GGCACGGGCACGTTCA (SEQ ID NO: 214). To normalize forvariations in loading and transfer efficiency membranes were strippedand probed for mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH)RNA (Clontech, Palo Alto, Calif.).

Example 21 Antisense Inhibition of Human HIF2α Expression by ChimericPhosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap

[0253] In accordance with the present invention, a series of antisensecompounds were designed to target different regions of the human HIF2αRNA, using published sequences (GenBank accession number NM_(—)001430.1,incorporated herein by reference and incorporated herein as SEQ ID NO:208). The compounds are shown in Table 5. “Target site” indicates thefirst (5′-most) nucleotide number on the particular target sequence towhich the compound binds. All compounds in Table 5 are chimericoligonucleotides (“gapmers”) 20 nucleotides in length, composed of acentral “gap” region consisting of ten 2′-deoxynucleotides, which isflanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”.The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. Theinternucleoside (backbone) linkages are phosphorothioate (P═S)throughout the oligonucleotide. All cytidine residues are5-methylcytidines. The compounds were analyzed for their effect on humanHIF2α mRNA levels by quantitative real-time PCR as described in otherexamples herein. Data are averages from three experiments in which A549cells were treated with the antisense oligonucleotides of the presentinvention. TABLE 5 Inhibition of human HIF2α mRNA levels by chimericphosphorothioate oligonucleotides having 2′-MOE wings and a deoxy gapTARGET SEQ ID TARGET % SEQ ID ISIS # REGION NO SITE SEQUENCE INHIB NO221985 Start 208 142 gtcagctgtcattgtcgctg 74 219 Codon 221987 Stop 2082751 ggcctggctcaggtggcctg 54 220 Codon 221989 Coding 208 1000ggtcatgttctcggagtcta 82 221 221991 Coding 208 1572 gtggagcagctgctgctgct80 222 221993 Coding 208 2412 ggtacatttgcgctcagtgg 76 223 221995 Coding208 2206 tgggcctcgagccccaaaac 15 224 221997 Coding 208 1300gaataggaagttactcttct 51 225 221999 Coding 208 1752 tggaagtcttccccgtccat69 226 222001 Coding 208 947 gcagctcctcagggtggtaa 82 227 222003 Coding208 977 catggtagaattcataggct 82 228 222005 Coding 208 1631tcacttcaatcttcaggtcg 55 229 222007 Coding 208 2691 gagcttcccagcacgggcac79 230 222009 Coding 208 1502 tgaaggcaggcaggctccca 77 231 222011 Coding208 2008 ggtgctggcctggccacagc 72 232 222013 Coding 208 561cgaatctcctcatggtcgca 89 233 222015 Coding 208 1247 tgctgttcatggccatcagg78 234 222017 Coding 208 1679 tactgcattggtccttggcc 78 235 222019 Coding208 1488 ctcccagcctcgctctgggt 63 236 222021 Coding 208 2700aggagcgtggagcttcccag 59 237 222023 Coding 208 623 ctgtggacatgtctttgctt79 238 222025 Coding 208 1716 agtgtctccaagtccagctc 84 239 222027 Coding208 759 ctattgtgaggagggcagtt 75 240 222029 Coding 208 237tcatagaacacctccgtctc 37 241 222031 Coding 208 2334 aaatgtgaggtgctgccacc67 242 222033 Coding 208 1578 ttgggcgtggagcagctgct 54 243 222035 Coding208 2126 gcgctgctcccaagaactct 89 244 222037 Coding 208 2639gcagcaggtaggactcaaat 64 245 222039 Coding 208 2325 gtgctgccaccaggtgggtc79 246 222041 Coding 208 1001 tggtcatgttctcggagtct 82 247 222043 Coding208 1209 tcagtctggtccatggagaa 80 248 222045 Coding 208 566tctcacgaatctcctcatgg 68 249 222047 Coding 208 1622 tcttcaggtcgttatccaaa56 250 222049 Coding 208 2715 aggtcccctccttgcaggag 66 251 222051 Coding208 246 tgggccagctcatagaacac 82 252 222053 Coding 208 2336tcaaatgtgaggtgctgcca 73 253 222055 Coding 208 391 catctgctggtcagcttcgg85 254 222057 Coding 208 1217 acagggattcagtctggtcc 84 255

[0254] As shown in Table 5, SEQ ID NOs 219, 220, 221, 211, 223, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254 and255 demonstrated at least 40% inhibition of HIF2α expression and aretherefore preferred. More preferred are SEQ ID NOs 233, 239 and 244. Thetarget regions to which these preferred sequences are complementary areherein referred to as “preferred target segments” and are thereforepreferred for targeting by compounds of the present invention. Thesepreferred target segments are shown in Table 7. These sequences areshown to contain thymine (T) but one of skill in the art will appreciatethat thymine (T) is generally replaced by uracil (U) in RNA sequences.The sequences represent the reverse complement of the preferredantisense compounds shown in Table 5. “Target site” indicates the first(5′-most) nucleotide number on the particular target nucleic acid towhich the oligonucleotide binds. Also shown in Table 7 is the species inwhich each of the preferred target segments was found.

Example 22 Antisense Inhibition of Mouse HIF2α Expression by ChimericPhosphorothioate Oligonucleotides Having 2′-MOE Wings and a Deoxy Gap

[0255] In accordance with the present invention, a second series ofantisense compounds were designed to target different regions of themouse HIF2α RNA, using published sequences (GenBank accession numberNM_(—)010137.1, incorporated herein by reference and incorporated hereinas SEQ ID NO: 212, nucleotides 20468925 to 20547619 of the sequence withGenBank accession number NW_(—)000133.1, incorporated herein byreference and incorporated herein as SEQ ID NO: 257, GenBank accessionnumber BY229956.1, incorporated herein by reference and incorporatedherein as SEQ ID NO: 258, and GenBank accession number AK087208.1,incorporated herein by reference and incorporated herein as SEQ ID NO:259). The compounds are shown in Table 6. “Target site” indicates thefirst (5′-most) nucleotide number on the particular target nucleic acidto which the compound binds. All compounds in Table 6 are chimericoligonucleotides (“gapmers”) 20 nucleotides in length, composed of acentral “gap” region consisting of ten 2′-deoxynucleotides, which isflanked on both sides (5′ and 3′-directions) by five-nucleotide “wings”.The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. Theinternucleoside (backbone) linkages are phosphorothioate (P═S)throughout the oligonucleotide. All cytidine residues are5-methylcytidines. The compounds were analyzed for their effect on mouseHIF2α mRNA levels by quantitative real-time PCR as described in otherexamples herein. Data are averages from three experiments in which b.ENDcells were treated with the antisense oligonucleotides of the presentinvention. TABLE 6 Inhibition of mouse HIF2α mRNA levels by chimericphosphorothioate oligonucleotides having 2′-MOE wings and a deoxy gapTARGET SEQ ID TARGET % SEQ ISIS # REGION NO: SITE SEQUENCE INHIB ID NO320972 5′UTR 212 130 ggttccttaaccccgtaggg 70 260 320973 5′UTR 212 135acctgggttccttaaccccg 61 261 320974 5′UTR 212 140 ggagcacctgggttccttaa 70262 320975 Start Codon 212 178 ttgtcagctgtcattgtcgc 72 263 320976 StartCodon 212 183 tctccttgtcagctgtcatt 84 264 320977 Coding 212 266gaagacctccgtctccttgc 83 265 320978 Coding 212 317 caggtgggagctcacactgt76 266 320979 Coding 212 352 aagctgatggccaggcgcat 64 267 320980 Coding212 442 ttcaggtacaagttatccat 78 268 320981 Coding 212 448aaggctttcaggtacaagtt 73 269 320982 Coding 212 461 aatgaaaccctccaaggctt87 270 320983 Coding 212 520 atgaacttgctgatgttttc 29 271 320984 Coding212 525 gtcccatgaacttgctgatg 57 272 320985 Coding 212 535acctgggtaagtcccatgaa 63 273 320986 Coding 212 545 tgttagttctacctgggtaa62 274 320987 Coding 212 563 gtcaaagatgctgtgtcctg 83 275 320988 Coding212 574 ggatgagtgaagtcaaagat 50 276 320989 Coding 212 673atgaagaagtcacgctcggt 63 277 320990 Coding 212 682 ttcatcctcatgaagaagtc53 278 320991 Coding 212 687 tgcacttcatcctcatgaag 58 279 320992 Coding212 714 tgacagtccggcctctgttg 52 280 320993 Coding 212 766actctcacttgcccggtgca 87 281 320994 Coding 212 776 gttgttgtagactctcactt64 282 320995 Coding 212 850 attggctcacacatgatgat 76 283 320996 Coding212 860 tgggtgctggattggctcac 75 284 320997 Coding 212 913atgctgtggcggctcaggaa 87 285 320998 Coding 212 970 gggtggtaaccaatcagttc76 286 320999 Coding 212 1057 gtgcacaagttctggtgact 50 287 321000 Coding212 1062 ccttggtgcacaagttctgg 74 288 321001 Coding 212 1135gtcccctgggtctccagcca 78 289 321002 Coding 212 1140 tgaccgtcccctgggtctcc63 290 321003 Coding 212 1145 gtagatgaccgtcccctggg 68 291 321004 Coding212 1150 gggttgtagatgaccgtccc 62 292 321005 Coding 212 1191catagttgacacacatgata 37 293 321006 Coding 212 1234 tccatggagaacaccacgtc76 294 321007 Coding 212 1239 tctggtccatggagaacacc 83 295 321008 Coding212 1286 aaagatgctgttcatggcca 51 296 321009 Coding 212 1338tggtgaacaggtagttgctc 64 297 321010 Coding 212 1363 agctcctcgggctcctcctt83 298 321011 Coding 212 1454 ggccttgccataggctgagg 49 299 321012 Coding212 1459 aggatggccttgccataggc 53 300 321013 Coding 212 1612ctgctgggcgtggagcagct 40 301 321014 Coding 212 1725 tgaagtccgtctgggtactg58 302 321015 Coding 212 1939 tccaactgctgcgggtactt 82 303 321016 Coding212 2002 ttgctcccagcatcaaagaa 0 304 321017 Coding 212 2012cagggaccctttgctcccag 81 305 321018 Coding 212 2038 gtgctggcctggccacagca66 306 321019 Coding 212 2216 cttgaacatggagacatgag 65 307 321020 Coding212 2226 cagacctcatcttgaacatg 72 308 321021 Coding 212 2231ctttgcagacctcatcttga 73 309 321022 Coding 212 2296 ttcagcttgttggacagggc51 310 321023 Coding 212 2376 gtgaactgctggtgcctgga 79 311 321024 Coding212 2386 cacatcaagtgtgaactgct 0 312 321025 Coding 212 2413ccgcccatgaggctcttcat 70 313 321026 Coding 212 2423 aggacaggtcccgcccatga85 314 321027 Coding 212 2433 caggcatcaaaggacaggtc 55 315 321028 Coding212 2482 gatttttgggtgaattcatc 38 316 321029 Coding 212 2647ctggccacgcctgacacctt 65 317 321030 Coding 212 2665 gatggccccagcagtcgact64 318 321031 Coding 212 2670 cgaacgatggccccagcagt 48 319 321032 Coding212 2680 aggtaaggctcgaacgatgg 65 320 321033 Coding 212 2707cagtcatatctggtcagttc 78 321 321034 Coding 212 2712 cctcacagtcatatctggtc83 322 321035 Coding 212 2717 gttcacctcacagtcatatc 66 323 321036 Coding212 2722 ggcacgttcacctcacagtc 81 324 321037 Coding 212 2727gcacgggcacgttcacctca 90 325 321038 Coding 212 2758 tctctcccctgcaggagtgt79 326 321039 Coding 212 2768 tctgagaaggtctctcccct 51 327 321040 Coding212 2778 ggtccagagctctgagaagg 73 328 321041 Stop Codon 212 2791gctcaggtggcctggtccag 69 329 321042 Stop Codon 212 2798ggccctggctcaggtggcct 12 330 321043 3′UTR 212 3199 agaacaagaacacttgagtt66 331 321044 intron 257 12633 aacagttgagacatgacagt 67 332 321045 exon:257 74580 tgtcactaacctcatcttga 45 333 intron junction 321046 5′UTR 258235 acaggagtcacttttctggg 43 334 321047 5′UTR 258 82 catacagtctcaggacactg47 335 321048 Genomic 259 116 aatctgtccatgaaaagaca 33 336

[0256] As shown in Table 6, SEQ ID NO, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 305, 306, 307, 308, 309, 310,311, 313, 314, 315, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326,327, 328, 329, 331, 332, 333, 334 and 335 demonstrated at least 40%inhibition of mouse HIF2α expression in this experiment and aretherefore preferred. More preferred are SEQ ID NOs 270, 281 and 285. Thetarget regions to which these preferred sequences are complementary areherein referred to as “preferred target segments” and are thereforepreferred for targeting by compounds of the present invention. Thesepreferred target segments are shown in Table 7. These sequences areshown to contain thymine (T) but one of skill in the art will appreciatethat thymine (T) is generally replaced by uracil (U) in RNA sequences.The sequences represent the reverse complement of the preferredantisense compounds shown in Tables 5 and 6. “Target site” indicates thefirst (5′-most) nucleotide number on the particular target nucleic acidto which the oligonucleotide binds. Also shown in Table 3 is the speciesin which each of the preferred target segments was found. TABLE 7Sequence and position of preferred target segments identified inhypoxia-inducible factor 2 alpha. TARGET SITE SEQ ID TARGET REV COMP SEQID ID NO SITE SEQUENCE OF SEQ ID ACTIVE IN NO 138730 208 142cagcgacaatgacagctgac 291 H. sapiens 337 138731 208 2751caggccacctgagccaggcc 292 H. sapiens 338 138732 208 1000tagactccgagaacatgacc 293 H. sapiens 339 138733 208 1572agcagcagcagctgctccac 294 H. sapiens 340 138734 208 2412ccactgagcgcaaatgtacc 295 H. sapiens 341 138736 208 1300agaagagtaacttcctattc 297 H. sapiens 342 138737 208 1752atggacggggaagacttcca 298 H. sapiens 343 138738 208 947ttaccaccctgaggagctgc 299 H. sapiens 344 138739 208 977agcctatgaattctaccatg 300 H. sapiens 345 138740 208 1631cgacctgaagattgaagtga 301 H. sapiens 346 138741 208 2691gtgcccgtgctgggaagctc 302 H. sapiens 347 138742 208 1502tgggagcctgcctgccttca 303 H. sapiens 348 138743 208 2008gctgtggccaggccagcacc 304 H. sapiens 349 138744 208 561tgcgaccatgaggagattcg 305 H. sapiens 350 138745 208 1247cctgatggccatgaacagca 306 H. sapiens 351 138746 208 1679ggccaaggaccaatgcagta 307 H. sapiens 352 138747 208 1488acccagagcgaggctgggag 308 H. sapiens 353 138748 208 2700ctgggaagctccacgctcct 309 H. sapiens 354 138749 208 623aagcaaagacatgtccacag 310 H. sapiens 355 138750 208 1716gagctggacttggagacact 311 H. sapiens 356 138751 208 759aactgccctcctcacaatag 312 H. sapiens 357 138753 208 2334ggtggcagcacctcacattt 314 H. sapiens 358 138754 208 1578agcagctgctccacgcccaa 315 H. sapiens 359 138755 208 2126agagttcttgggagcagcgc 316 H. sapiens 360 138756 208 2639atttgagtcctacctgctgc 317 H. sapiens 361 138757 208 2325gacccacctggtggcagcac 318 H. sapiens 362 138758 208 1001agactccgagaacatgacca 319 H. sapiens 363 138759 208 1209ttctccatggaccagactga 320 H. sapiens 364 138760 208 566ccatgaggagattcgtgaga 321 H. sapiens 365 138761 208 1622tttggataacgacctgaaga 322 H. sapiens 366 138762 208 2715ctcctgcaaggaggggacct 323 H. sapiens 367 138763 208 246gtgttctatgagctggccca 324 H. sapiens 368 138764 208 2336tggcagcacctcacatttga 325 H. sapiens 369 138765 208 391ccgaagctgaccagcagatg 326 H. sapiens 370 138766 208 1217ggaccagactgaatccctgt 327 H. sapiens 371 237138 212 130ccctacggggttaaggaacc 332 M. musculus 372 237139 212 135cggggttaaggaacccaggt 333 M. musculus 373 237140 212 140ttaaggaacccaggtgctcc 334 M. musculus 374 237141 212 178gcgacaatgacagctgacaa 335 M. musculus 375 237142 212 183aatgacagctgacaaggaga 336 M. musculus 376 237143 212 266gcaaggagacggaggtcttc 337 M. musculus 377 237144 212 317acagtgtgagctcccacctg 338 M. musculus 378 237145 212 352atgcgcctggccatcagctt 339 M. musculus 379 237146 212 442atggataacttgtacctgaa 340 M. musculus 380 237147 212 448aacttgtacctgaaagcctt 341 M. musculus 381 237148 212 461aagccttggagggtttcatt 342 M. musculus 382 237150 212 525catcagcaagttcatgggac 344 M. musculus 383 237151 212 535ttcatgggacttacccaggt 345 M. musculus 384 237152 212 545ttacccaggtagaactaaca 346 M. musculus 385 237153 212 563caggacacagcatctttgac 347 M. musculus 386 237154 212 574atctttgacttcactcatcc 348 M. musculus 387 237155 212 673accgagcgtgacttcttcat 349 M. musculus 388 237156 212 682gacttcttcatgaggatgaa 350 M. musculus 389 237157 212 687cttcatgaggatgaagtgca 351 M. musculus 390 237158 212 714caacagaggccggactgtca 352 M. musculus 391 237159 212 766tgcaccgggcaagtgagagt 353 M. musculus 392 237160 212 776aagtgagagtctacaacaac 354 M. musculus 393 237161 212 850atcatcatgtgtgagccaat 355 M. musculus 394 237162 212 860gtgagccaatccagcaccca 356 M. musculus 395 237163 212 913ttcctgagccgccacagcat 357 M. musculus 396 237164 212 970gaactgattggttaccaccc 358 M. musculus 397 237165 212 1057agtcaccagaacttgtgcac 359 M. musculus 398 237166 212 1062ccagaacttgtgcaccaagg 360 M. musculus 399 237167 212 1135tggctggagacccaggggac 361 M. musculus 400 237168 212 1140ggagacccaggggacggtca 362 M. musculus 401 237169 212 1145cccaggggacggtcatctac 363 M. musculus 402 237170 212 1150gggacggtcatctacaaccc 364 M. musculus 403 237172 212 1234gacgtggtgttctccatgga 366 M. musculus 404 237173 212 1239ggtgttctccatggaccaga 367 M. musculus 405 237174 212 1286tggccatgaacagcatcttt 368 M. musculus 406 237175 212 1338gagcaactacctgttcacca 369 M. musculus 407 237176 212 1363aaggaggagcccgaggagct 370 M. musculus 408 237177 212 1454cctcagcctatggcaaggcc 371 M. musculus 409 237178 212 1459gcctatggcaaggccatcct 372 M. musculus 410 237179 212 1612agctgctccacgcccagcag 373 M. musculus 411 237180 212 1725cagtacccagacggacttca 374 M. musculus 412 237181 212 1939aagtacccgcagcagttgga 375 M. musculus 413 237183 212 2012ctgggagcaaagggtccctg 377 M. musculus 414 237184 212 2038tgctgtggccaggccagcac 378 M. musculus 415 237185 212 2216ctcatgtctccatgttcaag 379 M. musculus 416 237186 212 2226catgttcaagatgaggtctg 380 M. musculus 417 237187 212 2231tcaagatgaggtctgcaaag 381 M. musculus 418 237188 212 2296gccctgtccaacaagctgaa 382 M. musculus 419 237189 212 2376tccaggcaccagcagttcac 383 M. musculus 420 237191 212 2413atgaagagcctcatgggcgg 385 M. musculus 421 237192 212 2423tcatgggcgggacctgtcct 386 M. musculus 422 237193 212 2433gacctgtcctttgatgcctg 387 M. musculus 423 237195 212 2647aaggtgtcaggcgtggccag 389 M. musculus 424 237196 212 2665agtcgactgctggggccatc 390 M. musculus 425 237197 212 2670actgctggggccatcgttcg 391 M. musculus 426 237198 212 2680ccatcgttcgagccttacct 392 M. musculus 427 237199 212 2707gaactgaccagatatgactg 393 M. musculus 428 237200 212 2712gaccagatatgactgtgagg 394 M. musculus 429 237201 212 2717gatatgactgtgaggtgaac 395 M. musculus 430 237202 212 2722gactgtgaggtgaacgtgcc 396 M. musculus 431 237203 212 2727tgaggtgaacgtgcccgtgc 397 M. musculus 432 237204 212 2758acactcctgcaggggagaga 398 M. musculus 433 237205 212 2768aggggagagaccttctcaga 399 M. musculus 434 237206 212 2778ccttctcagagctctggacc 400 M. musculus 435 237207 212 2791ctggaccaggccacctgagc 401 M. musculus 436 237209 212 3199aactcaagtgttcttgttct 403 M. musculus 437 237210 257 12633actgtcatgtctcaactgtt 404 M. musculus 438 237211 257 74580tcaagatgaggttagtgaca 405 M. musculus 439 237212 258 235cccagaaaagtgactcctgt 406 M. musculus 440 237213 258 82cagtgtcctgagactgtatg 407 M. musculus 441

[0257] As these “preferred target segments” have been found byexperimentation to be open to, and accessible for, hybridization withthe antisense compounds of the present invention, one of skill in theart will recognize or be able to ascertain, using no more than routineexperimentation, further embodiments of the invention that encompassother compounds that specifically hybridize to these preferred targetsegments and consequently inhibit the expression of HIF2α.

[0258] According to the present invention, antisense compounds includeantisense oligomeric compounds, antisense oligonucleotides, ribozymes,external guide sequence (EGS) oligonucleotides, alternate splicers,primers, probes, and other short oligomeric compounds which hybridize toat least a portion of the target nucleic acid.

Example 23 Expression of HIF1α and HIF2α in Various Human Cell Lines

[0259] U87-MG human glioblastoma, PC-3 human prostate cancer, JEG-3human choriocarcinoma, HeLa human cervix cancer, SK-N-BE(2)neuroblastoma, MCF-7 human breast cancer, 786-O human clear-cell renalcell carcinoma, Calu-1 human lung cancer, and Hep3B human hepatocellularcarcinoma cells were purchased from American Type Culture Collection(ATCC; Manassas, Va.) and cultured according to ATCC directions. Humanumbilical endothelial cells (HUVEC) were obtained from Cascade Biologics(Portland Oreg.). Hypoxic treatments of cells (0.5-0.8×10⁶/60 mm dish or1-2×10⁶/100 mm dish) were performed at 1% O₂ in a chamber controlled byProOx oxygen sensor (BioSpherix, Redfield, N.Y.) for 16 h. To achievethe optimal hypoxic induction, 3 or 6 ml of medium was used for 60 mmand 100 mm dish culture, respectively during incubation. CoCl₂ (150 μM)was added to the cells to mimic hypoxic condition in some experiments.

[0260] Cultured cells at normoxia, hypoxia, or with CoCl₂ were harvestedand whole cell lysates prepared with RIPA buffer containing proteaseinhibitor cocktails (Roche), 0.5 mM sodium orthovanadate, 10 mMβ-glycerophophate, 250 ng/ml ubiquitin aldehyde, and 400 nM epoxomicinwere separated on 12% SDS-PAGE and transferred to PVDF membranes(Amersham Biosciences). Typically, 35-50 μg of proteins were loaded perlane. Immunoblotting was performed with the following antibodies:anti-HIF-1α (BD Transduction Laboratories) at 1:250 (v/v); anti-HIF-2α(EPAS1) (Santa Cruz Biotechnology Inc) at 1:150; anti-HIF-1β (BDTransduction Laboratories) at 1:1000; anti-VHL (BD TransductionLaboratories) at 1:500; anti-GLUT-1 (Alpha Diagnostic International) at1:600, and anti-α-tubulin (Sigma) at 1:2000 in 0.05%Tween-20/Tris-buffered saline (T-TBS) blocking buffer containing 5%nonfat skim milk at 4° C. overnight, followed by washing with T-TBS for30 min. Goat anti-mouse or rabbit IgGs coupled with HRP (BioRad) wereused as secondary antibodies at 1:3000. Immunospecific bands weredetected by enhanced chemiluminescence plus (ECL-Plus) detection kit(Amersham Biosciences).

[0261] Hif1α expression was shown to be increased in hypoxic conditionsand in the presence of CoCl₂ (which mimics hypoxia) in U87-MG humanglioblastoma, PC-3 human prostate cancer, JEG-3 human choriocarcinoma,HeLa human cervix cancer, SK-N-BE(2) neuroblastoma, MCF-7 human breastcancer, Calu-1 human lung cancer, and Hep3B human hepatocellularcarcinoma cells but not 786-O human clear-cell renal cell carcinomacells.

[0262] Hif2α expression was shown to be increased in hypoxic (1% O₂)conditions and in the presence of CoCl₂ in U87-MG human glioblastoma,PC-3 human prostate cancer, JEG-3 human choriocarcinoma, MCF-7 humanbreast cancer, 786-O human clear-cell renal cell carcinoma, Calu-1 humanlung cancer, Hep3B human hepatocellular carcinoma cells and HUVECs.

Example 24 Antisense Modulation of HIF1α mRNA Expression in Cancer Cells(Dose Response)

[0263] HeLa, Hep3B, or U87-MG cells were plated in 96-well plates(8-10,000/well) 16 h prior to transfection. The following antisenseoligonucleotides were delivered into cells by lipofectin (3 μg/ml per100 nM oligonucleotide) in Opti-Mem media (Invitrogen) at the indicatedconcentration: ISIS 175510 (SEQ ID NO: 47) and ISIS 298697 (SEQ ID NO:141) are targeted to human HIF-1α ASOs; ISIS 222035 (SEQ ID NO: 244) istargeted to human HIF-2α; and ISIS 129688 (TTCGCGGCTGGACGATTCAG; SEQ IDNO: 442) is an unrelated control. 10/35 is an equal mixture of ISIS175510 and 222035 (HIF1α and HIF2α inhibitory oligonucleotides). ISIS97/35 is an equal mixture of ISIS 298697 and ISIS 222035 (HIF1α andHIF2α inhibitory oligonucleotides).

[0264] The transfection medium was switched to complete growth medium(120 μl/well) 4 h after transfection. Sixty microliters of medium wasremoved from the well 3 h after media switch and the cells were furtherincubated at normoxia or hypoxia for 16-20 h. TABLE 8 HIF1α mRNAexpression in antisense treated HeLa cells Shown as percent inhibitionrelative to control oligonucleotide N = Normoxia (21% O₂) H = Hypoxia(1% O₂) Percent inhibition of HIF1α mRNA Oligo- Normoxia expressionafter treatment with nucleotide or oligonucleotide at concentrationsshown: and conditions: Hypoxia 0 6.25 nM 25 nM 100 nM 200 nM 129688 N 00 0 30 47 129688 H 1 1 10 30 57 175510 N 0 24 77 94 94 175510 H 1 39 8295 96 298697 N 0 44 72 91 93 298697 H 3 30 75 92 93 222035 N 0 0 0 1 24222035 H 3 3 0 11 35 10/35 N 0 33 82 94 94 10/35 H 3 35 85 94 95 97/35 N0 16 66 84 85 97/35 H 3 34 79 88 89

[0265] It can be seen that the HIF1α antisense oligonucleotides ISIS175510 and 298697 specifically inhibited HIF1α and not HIF2α. Similarresults were obtained in Hep3b human hepatocellular carcinoma cells andin U87-MG human glioblastoma cells.

Example 25 Antisense Modulation of HIF2α mRNA expression in cancer cells(Dose Response)

[0266] HeLa, Hep3B, or U87-MG cells were plated in 96-well plates(8-10,000/well) 16 h prior to transfection. The following antisenseoligonucleotides were delivered into cells by lipofectin (3 μg/ml per100 nM oligonucleotide) in Opti-Mem media (Invitrogen) at the indicatedconcentration: ISIS (SEQ ID NO: 47) and ISIS 298697 (SEQ ID NO: 141) aretargeted to human HIF-1α ASOs; ISIS 222035 (SEQ ID NO: 244) is targetedto human HIF-2α; and ISIS 129688 (TTCGCGGCTGGACGATTCAG; SEQ ID NO: 442)is an unrelated control. 10/35 is an equal mixture of ISIS 175510 and222035 (HIF1α and HIF2α inhibitory oligonucleotides). ISIS 97/35 is anequal mixture of ISIS 298697 and ISIS 222035 (HIF1α and HIF2α inhibitoryoligonucleotides). The transfection medium was switched to completegrowth medium (120 μl/well) 4 h after transfection. Sixty microliters ofmedium was removed from the well 3 h after media switch and the cellswere further incubated at normoxia or hypoxia for 16-20 h. TABLE 9 HIF2αmRNA expression in ASO treated HeLa cells Shown as percent inhibitionrelative to control oligonucleotide N = Normoxia (21% O₂) H = Hypoxia(1% O₂) Percent inhibition of HIF1α mPNA Oligo- Normoxia expressionafter treatment with nucleotide or oligonucleotide at concentrationsshown: and conditions: Hypoxia 0 6.25 nM 25 nM 100 nM 200 nM 129688 N 00 16 12 21 129688 H 0 0 4 12 50 175510 N 0 1 0 0 0 175510 H 0 8 0 4 0298697 N 0 0 10 48 65 298697 H 0 0 11 52 58 222035 N 0 0 62 93 96 222035H 0 19 73 94 96 10/35 N 0 0 77 96 96 10/35 H 0 21 78 94 95 97/35 N 0 063 89 95 97/35 H 0 34 79 96 96

[0267] It can be seen that the HIF2α antisense oligonucleotide ISIS222035 specifically inhibited HIF2α relative to HIF1α. Theoligonucleotide ISIS 298697, designed to target human HIF1α, showed someability to inhibit HIF2α expression as well. This oligonucleotide hasperfect complementarity to the HIF1α target sequence and was found tohave only two mismatches to the human HIF2α. Similar results were seenin U87-MG human glioblastoma cells and HepG3 hepatocellular carcinomacells.

Example 26 HIF2α Plays a Major Role in the Induction of VEGF by Hypoxiain U87-MG Cells

[0268] Genes whose products are dramatically induced by hypoxia (orCoCl₂, a mimic of hypoxia) include erythropoietin (Epo), glucosetransporter-1 (Glut-1), vascular endothelial growth factor (VEGF) andPhosphofructokinase-L (PFK-L). They are induced by hypoxia to varyingextents in various cell lines. As shown in previous examples, VEGFexpression is induced by hypoxia in U87-MG cells. The followingantisense oligonucleotides were delivered into cells by lipofectin (3μg/ml per 100 nM oligonucleotide) in Opti-Mem media (Invitrogen) at theindicated concentration: ISIS 175510 (SEQ ID NO: 47) and ISIS 298697(SEQ ID NO: 141) are targeted to human HIF-1α ASOs; ISIS 222035 (SEQ IDNO: 244) is targeted to human HIF-2α; and ISIS 129688(TTCGCGGCTGGACGATTCAG; SEQ ID NO: 442) is an unrelated control. 10/35 isan equal mixture of ISIS 175510 and 222035 (HIF1α and HIF2α inhibitoryoligonucleotides). ISIS 97/35 is an equal mixture of ISIS 298697 andISIS 222035 (HIF1α and HIF2α inhibitory oligonucleotides). TABLE 10HIF2α plays a major role in the induction of VEGF by hypoxia in U87-MGcells Oligo- Relative VEGF nucleotide Normoxia mRNA expression aftertreatment with and or oligonucleotide at concentrations shown:conditions: Hypoxia 0 6.25 nM 25 nM 100 nM 200 nM 129688 N 100 103 11173 81 129688 H 372 378 346 363 383 175510 N 100 86 65 61 62 175510 H 372397 407 338 392 298697 N 100 111 81 56 73 298697 H 372 413 342 312 275222035 N 100 94 69 48 45 222035 H 372 399 257 131 108 10/35 N 100 81 4845 44 10/35 H 372 431 254 110 80 97/35 N 100 119 63 45 47 97/35 H 372409 289 124 85

[0269] ISIS 175510, which specifically inhibits HIF1α and not HIF2α, wasfound to have no effect on VEGF induction by hypoxia in U87-MG cells. Incontrast, ISIS 222035, which specifically inhibits HIF2α and not HIF1α,caused a dose-dependent decrease in VEGF induction. ISIS 298697, whichwas designed to target HIF1α but was found to have crossreactivity withHIF2α, also interfered with VEGF induction by hypoxia. Thus HIF2α playsa major role in the induction of VEGF by hypoxia in U87-MG cells.

Example 27 HIF2α Plays a Major Role in the Induction of Epo by Hypoxiain Hep3B Cells

[0270] Genes whose products are dramatically induced by hypoxia (orCoCl₂, a mimic of hypoxia) include Epo, Glut-1, VEGF and PFK-L. They areinduced by hypoxia to varying extents in various cell lines. Epo(erythropoietin) expression is induced by hypoxia in Hep3B cells. Thefollowing antisense oligonucleotides were delivered into Hep3B cells bylipofectin (3 μg/ml per 100 nM oligonucleotide) in Opti-Mem media(Invitrogen) at the indicated concentration: ISIS 175510 (SEQ ID NO: 47)and ISIS 298697 (SEQ ID NO: 141) are targeted to human HIF-1α ASOs; ISIS222035 (SEQ ID NO: 244) is targeted to human HIF-2α; and ISIS 129688(TTCGCGGCTGGACGATTCAG; SEQ ID NO: 442) is an unrelated control. 10/35 isan equal mixture of ISIS 175510 and 222035 (HIF1α and HIF2α inhibitoryoligonucleotides). ISIS 97/35 is an equal mixture of ISIS 298697 andISIS 222035 (HIF1α HIF2α inhibitory oligonucleotides). TABLE 11 HIF2αplays a major role in the induction of Epo by hypoxia in Hep3B cellsRelative EPO mRNA expression after treatment with oligonucleotide atconcentrations shown: Normoxia Shown as -Fold induction over controloligonucleotide or 6.25 and conditions: Hypoxia 0 nM 25 nM 100 nM 200 nM129688 N 1 1 0 3 11 129688 H 531 586 433 261 128 175510 N 1 8 3 3 2175510 H 531 577 542 326 144 298697 N 1 9 11 12 3 298697 H 531 436 32652 6 222035 N 1 3 3 2 1 222035 H 531 302 101 2 2 10/35 N 1 2 0 0 3 10/35H 531 212 30 0 1 97/35 N 1 2 0 1 4 97/35 H 531 194 29 2 1

[0271] ISIS 175510, which specifically inhibits HIF1• and not HIF2•, wasfound to have no effect on Epo induction by hypoxia in Hep3B cells. Incontrast, ISIS 222035, which specifically inhibits HIF2• and not HIF1•,caused a dose-dependent decrease in Epo induction. ISIS 298697, whichwas designed to target HIF1• but was found to have crossreactivity withHIF2•, also interfered with Epo induction by hypoxia. Thus HIF2a plays amajor role in the induction of Epo by hypoxia in Hep3B cells.

Example 28 Both HIF1α and HIF2α Play a Major Role in the Induction ofVEGF by Hypoxia in HeLa Cells

[0272] Genes whose products are dramatically induced by hypoxia (orCoCl₂) include Epo (erythropoietin), Glut-1, VEGF andPhosphofructokinase (PFK)-L. They are induced by hypoxia to varyingextents in various cell lines. VEGF expression is induced by hypoxia inHeLa cells. The following antisense oligonucleotides were delivered intocells by lipofectin (3 μg/ml per 100 nM oligonucleotide) in Opti-Memmedia (Invitrogen) at the indicated concentration: ISIS 175510 (SEQ IDNO: 47) and ISIS 298697 (SEQ ID NO: 141) are targeted to human HIF-1αASOs; ISIS 222035 (SEQ ID NO: 244) is targeted to human HIF-2α; and ISIS129688 (TTCGCGGCTGGATTCAG; SEQ ID NO: 442) is an unrelated control.10/35 is an equal mixture of ISIS 175510 and 222035 (HIF1• and HIF2•inhibitory oligonucleotides). ISIS 97/35 is an equal mixture of ISIS298697 and ISIS 222035 (HIF1• and HIF2• inhibitory oligonucleotides).TABLE 12 HIF1α and HIF2α play a major role in the induction of VEGF byhypoxia in HeLa cells Oligo- Relative nucleotide Normoxia VEGF mRNAexpression after treatment and or with oligonucleotide at concentrationsshown: conditions: Hypoxia 0 6.25 nM 25 nM 100 nM 200 nM 129688 N 100119 100 85 93 129688 H 284 283 289 234 209 175510 N 100 95 132 110 113175510 H 284 249 157 113 106 298697 N 100 84 105 93 93 298697 H 284 211144 106 108 222035 N 100 111 114 92 67 222035 H 284 260 209 144 77 10/35N 100 94 97 76 58 10/35 H 284 214 104 74 70 97/35 N 100 106 80 65 5697/35 H 284 207 108 73 60

[0273] In this experiment all oligonucleotides except for the control(129688) interfered with induction of VEGF by hypoxia in HeLa cells.Thus both HIF1α and HIF2α play a major role in the induction of VEGF byhypoxia in HeLa cells. Because the relative role of HIF1α and HIF2α inhypoxic induction depends both on cell type and by induced gene (e.g.,VEGF vs Epo), it is believed to be preferred to target both HIF1α andHIF2α for antisense inhibition. This may be achieved by a singlecross-HIF antisense compound (such as ISIS 298697) or by a combinationof one or more antisense compounds targeted to HIF1α and one or moreantisense compounds targeted to HIF2α. Compounds administered incombination may be given simultaneously or sequentially.

Example 29 Designing and Testing HIF1•/HIF2• Cross-Reacting AntisenseCompounds

[0274] The human HIF1α and HIF2α target sequences were compared forregions of identity but none were found to be as long as 20 nucleotides.However, based on the somewhat limited sequence homology between thehuman HIF1α and HIF2α target sequences, a series of antisense sequenceswere designed which were perfectly complementary to either HIF1α orHIF2α and which had no more than 4 mismatches to the other HIFα. Thesecompounds are shown in Table 13. The primary target sequence (perfectcomplementarity) is shown in the “target” column and the number ofmismatches against the other HIF is shown in subsequent columns. “Targetsite” refers to position on the primary target sequence. TABLE 13HIF1α/HIF2α crossreacting antisense sequences SEQ #Mis- # Mis- ID matchvs matches vs Target ISIS NO OLIGO_SEQ NO Target HIF1α HIF2α siteHIF1α˜EC50 HIF2α˜EC50 129688 TTCGCGGCTGGACGATTCAG 442 Control 330460CCTCATGGTCGCAGGGATGA 443 HIF2α 2 557 (G-A, G-U) 330462TCTCCTCATGGTCGCAGGGA 444 HIF2α 3 557 (G-A, G-U, C-A) 222013CGAATCTCCTCATGGTCGCA 233 HIF2α 4 561 (G-U, C-A, A-G, G-A) 298697TCCTCATGGTCACATGGATG 141 HIF1α 2 673 5 30 (A-C, T-C) 330447TCATGGTCACATGGATGAGT 445 HIF1α 2 670 8 50 A-C, T-C) 330449CCTCATGGTCACATGGATGA 446 HIF1α 2 672 5 30 A-C, T-C) 330448CTCATGGTCACATGGATGAG 447 HIF1α 2 671 (A-C, T-C) 330452ATTTCCTCATGGTCACATGG 448 HIF1α 3 676 (A-C, T-C, G-T) 330470AAACCCTCCAAGGCTTTCAG 449 HIF2α 2 423 45 20 (G-U, C-U) 326743TCCTCATGGTCGCAGGGATG 450 HIF2α 2 555 40 10 G-A, G-U)

[0275] Thus it is possible to inhibit both HIF1α and HIF2α with a singlecrossreacting oligonucleotide, although the relative antisense efficacyis not equal for the two forms because of imperfect homology to one HIFαor the other.

Example 30 Crossr Acting HIF1α/HIF2α Antisense Compounds Containing“Universal” Bases

[0276] In order to try to get antisense compounds that were highlypotent against both HIF1α and HIF2α targets, the nucleobases at thesites of the mismatches against one or the other HIF were replaced withthe “universal bases” inosine or 3′nitro-pyrrole. Inosine has theability to pair with G, U or C. If there was an A at the particularposition of either of the sequences, we used 3-nitropyrrole. This is abase that does not have binding affinity to any of the bases, but alsodoes not cause steric hindrance of the duplex. These oligos werescreened and found to be active against both targets with anintermediate potency. This is shown in Table 14. In the table, “I”indicates inosine and “P” indicates 3-nitropyrrole. TABLE 14 HIF1α/HIF2αcrossreacting antisense compounds containing universal bases SEQ # Mis-# Mis- ID match vs match vs Target ISIS NO OLIGO_SEQ NO Target HIF1αHIF2α site HIF1α˜EC50 HIF2α˜EC50 326743 TCCTCATGGTCGCAGGGATG 450 HIF2α 2555 40 10 298697 TCCTCATGGTCACATGGATG 141 HIF1α 2 673 5 30 (A-C, T-C)330449 CCTCATGGTCAPCATGGATGA 446 HIF1α 2 672 5 30 (A-C, T-C) 337223TCCTCATGGTCICAPGGATG 451 HIF1α 2 2 673 25 15 and (I-T, P-A) (I-C, P-C)HIF2α 337224 CCTCATGGTCICAPGGATGA 452 HIF1α 2 2 672 25 15 and (I-T, P-A)HIF2α

[0277] Introduction of universal bases into the antisense compounds atthe site of mismatches resulted in a more equal inhibitory effect forboth HIF1α and HIF2α.

Example 31 Tube Formation Assay to Determine Effect of HIF1α and HIF2αAntisense Inhibitors on Angiogenesis

[0278] Angiogenesis is stimulated by numerous factors that promoteinteraction of endothelial cells with each other and with extracellularmatrix molecules, resulting in the formation of capillary tubes. Thisprocess can be reproduced in tissue culture by the formation oftube-like structures by endothelial cells. Loss of tube formation invitro has been correlated with the inhibition of angiogenesis in vivo(Carmeliet et al., (2000) Nature 407:249-257; and Zhang et al., (2002)Cancer Research 62:2034-42), which supports the use of in vitro tubeformation as an endpoint for angiogenesis.

[0279] Angiogenesis, or neovascularization, is the formation of newcapillaries from existing blood vessels. In adult organisms this processis typically controlled and short-lived, for example in wound repair andregeneration. However, aberrant capillary growth can occur and thisuncontrolled growth plays a causal and/or supportive role in manypathologic conditions such as tumor growth and metastasis. In thecontext of this invention “aberrant angiogenesis” refers to unwanted oruncontrolled angiogenesis. Angiogenesis inhibitors are being evaluatedfor use as antitumor drugs. Other diseases and conditions associatedwith angiogenesis include arthritis, cardiovascular diseases, skinconditions, and aberrant wound healing. Aberrant angiogenesis can alsooccur in the eye, causing loss of vision. Examples of ocular conditionsinvolving aberrant angiogenesis include macular degeneration, diabeticretinopathy and retinopathy of prematurity.

[0280] The tube formation assay is performed using an in vitroAngiogenesis Assay Kit (Chemicon International, Temecula, Calif.), orgrowth factor reduced Matrigel (BD Biosciences, Bedford, Mass.). HUVECswere plated at 4000 cells/well in 96-well plates. One day later, cellswere transfected with antisense and control oligonucleotides accordingto standard published procedures (Monia et al., (1993) J Biol Chem. 1993Jul. 5;268(19):14514-22) using 75 nM oligonucleotide in lipofectin(Gibco, Grand Island, N.Y.). Approximately fifty hourspost-transfection, cells were transferred to 96-well plates coated withECMatrix™ (Chemicon International) or growth factor depleted Matrigel.Under these conditions, untreated HUVECs form tube-like structures.After an overnight incubation at 37° C., treated and untreated cellswere inspected by light microscopy. Individual wells were assigneddiscrete scores from 1 to 5 depending on the extent of tube formation. Ascore of 1 refers to a well with no tube formation while a score of 5 isgiven to wells where all cells are forming an extensive tubular network.

Table 15 Effect of HIF1α and HIF2α Antisense Oligonucleotides onAngiogenic Tube Formation

[0281] ISIS 29848 (NNNNNNNNNNNNNNNNNNNN; SEQ ID NO: 453) is a controloligonucleotide containing an equal mixture of the bases A, C, G and Tat every position. ISIS 298695 (SEQ ID NO: 139) and ISIS 298750 (Seq;SEQ ID NO: 193) are targeted to HIF1α; ISIS 330447 (Seq; SEQ ID NO: 445)is a cross-HIF oligonucleotide having perfect complementarity to HIF1αtarget and imperfect complementarity (and thus less inhibitory effect)for HIF2α; ISIS 222035 (SEQ ID NO: 244) and 222025 (SEQ ID NO: 239) aretargeted to HIF2α and ISIS 326743 is a cross-HIF oligonucleotide havingperfect complementarity to HIF2α target and imperfect complementarity(and thus less inhibitory effect) for HIF1α. ISIS # Target 0 10 nM 35 nM75 nM  29848 control 5 5 4.75 4.375 298695 HIF1α 5 5 5 3.75 298750 HIF1α5 5 4.75 3.25 330447 HIF 1α/ 5 5 4.25 3 2α 222035 HIF2α 5 5 3.75 1.75222025 HIF2α 5 5 3.5 1.75 326743 HIF2α/ 5 5 4.75 5 1α

[0282] As calculated from the assigned discrete scores, it is apparentthat HUVEC tube formation is inhibited by reduction of HIF2α and HIF1α,singly or in combination.

Example 32 Inhibition of HIF1α Expression In Vivo

[0283] C57B1/6 mice are maintained on a standard rodent diet and areused as control (lean) animals. Seven-week old male C57B1/6 mice areinjected subcutaneously with oligonucleotides at a dose of 25 mg/kg twotimes per week for 4 weeks. Saline-injected animals serve as a control.After the treatment period, mice are sacrificed and target levels areevaluated in liver, using RNA isolation and target mRNA expression levelquantitation (RT-PCR) as described in other examples herein.

[0284] Oligonucleotides used in this experiment were ISIS 298695 (SEQ IDNO: 139), ISIS 298697 (SEQ ID NO: 141), and ISIS 298750, (SEQ ID NO:193), all targeted to mouse HIF1• and crossreactive to human HIF1• ISIS141923 (CCTTCCCTGAAGGTTCCTCC; SEQ ID NO: 454) is an unrelated negativecontrol oligonucleotide. Results are shown in Table 16. TABLE 16Antisense inhibition of HIF1• expression in mouse liver by antisense toHIF1• ISIS # % inhib. of HIF1• Saline 0 ISIS 298695 76 ISIS 298697 70ISIS 298750 74 ISIS 141923 0 (control)

[0285] The effect of inhibiting HIF1α on levels of VEGF and GLUT1 inmouse liver was also determined. These are both hypoxia-inducibletargets. Results are shown in Table 17 and 18. TABLE 17 Effect ofAntisense inhibition of HIF1• on VEGF expression in mouse liver ISIS # %inhib. of VEGF Saline 0 ISIS 298695 12 ISIS 298697 4 ISIS 298750 0 ISIS141923 0 (control)

[0286] TABLE 18 Effect of antisense inhibition of HIF1• on GLUT1expression in mouse liver ISIS # % inhib. of VEGF Saline 0 ISIS 298695 0ISIS 298697 15 ISIS 298750 0 ISIS 141923 22 (control)

Example 33 Antisense Inhibition of HIF1α in a Mouse Model ofHepatocellular Carcinoma (HCC)

[0287] An HCC mouse model (C57BL/6-TgN(CRP-TagSV40)60-4, Taconic,Germantown N.Y.) for hepatocellular carcinoma was used in whichtransgenic male mice express SV40 T-antigen (Tag) in their livers, whichleads to spontaneous development of well-differentiated hepatocellularcarcinoma (HCC) carcinomas (Ruther et al., 1993, Oncogene 8, 87-93). HCCmice were treated with ISIS 298695, ISIS 298697 or ISIS 298750, alltargeted to HIF1• or with an unrelated control oligonucleotide. HCC andwild type mice were also treated with saline alone. Results are shown inTable 19. TABLE 19 Antisense inhibition of HIF1• in HCC mouse liver ISIS# SEQ ID NO % inhib. of HIF1• Saline 0 ISIS 298695 139 43 ISIS 298697141 33 ISIS 298750 193 40 ISIS 141923 454 11 (control) C57BL6/saline 43

[0288] The effect of HIF1• inhibition on GLUT1 expression in HCC micewas also evaluated. Results are shown in Table 20. TABLE 20 Effect ofantisense inhibition of HIF1• on GLUT1 levels in HCC mouse liver ISIS #SEQ ID NO % inhib. of GLUT1 Saline 0 ISIS 298695 139 0 ISIS 298697 141 0ISIS 298750 193 13 ISIS 141923 454 18 (control) C57BL6/saline 2

Example 34 Inhibition of HIF2α Expression in Tumor Cells by Wild-Typep53 Under Hypoxia in T47D Tumor Cells

[0289] T47D breast adenocarcinoma cells were obtained from the AmericanType Culture Collection (ATCC) (Manassas, Va.). Cells were cultured inGibco DMEM High glucose media supplemented with 10% FBS. p53 is a tumorsuppressor gene product which is inactive or aberrant in approximately50% of human tumors. T47D cells are p53 null, i.e. they contain inactivemutant p53. These cells express high levels of HIF2• even in normoxicconditions. Hypoxia or CoCl₂ induces even higher levels of HIF2•expression. In contrast, T47D cells which have been transfected with aplasmid expressing p53, thus restoring p53 function in these cells,express extremely low levels of HIF2•, even in hypoxic conditions or inCoCl₂ simulation of hypoxia. This increase in HIF2• in cells withaberrant p53 is believed to be a novel observation and is believed toindicate a link between p53 and the HIF pathway.

Example 35 Effects of Antisense Inhibition of HIF1α and/or HIF2α onCancer Cell Proliferation Under Hypoxia/Glucose Deprivation

[0290] PC-3 human prostate cancer cells were cultured as described inprevious examples. Cells were electroporated with oligonucleotide atconcentrations described below and grown for 16 hours at normoxia and0.45 g/l glucose. The medium was then replaced with either glucose (4.5g/l glucose) or low-glucose medium (no added glucose) and cells werethen kept at hypoxia (1% O₂) or normoxia (21% O₂) for another 48 hours.The effect of antisense treatment on cell proliferation was measured.Oligonucleotides were ISIS 129688 (unrelated control), ISIS 175510(HIF1α), ISIS 222035 (HIF2α) and ISIS 298697 (HIF1α with somecrossreactivity to HIF2α). Results are shown in the tables below, onetable for each culture condition. TABLE 21 Effect of HIF antisense onproliferation of PC-3 cancer cells Normoxia/Glucose Cell proliferationas percent of saline control ISIS # 0 nM 10 nM 20 nM SEQ ID NO 129688100 103 103 442 175510 100 126 93 47 222035 100 130 116 244 298697 100118 86 141

[0291] TABLE 22 Effect of HIF antisense on proliferation of PC-3 cancercells Hypoxia/Glucose Cell proliferation as percent of saline controlISIS # 0 nM 10 nM 20 nM SEQ ID NO 129688 100 104 99 442 175510 100 113105 47 222035 100 106 91 244 298697 100 113 83 141

[0292] TABLE 23 Effect of HIF antisense on proliferation of PC-3 cancercells Normoxia/Low Glucose Cell proliferation as percent of salinecontrol ISIS # 0 nM 10 nM 20 nM SEQ ID NO 129688 100 107 105 442 175510100 96 89 47 222035 100 91 68 244 298697 100 91 88 141

[0293] TABLE 24 Effect of HIF antisense on proliferation of PC-3 cancercells Hypoxia/Low Glucose Cell proliferation as percent of salinecontrol ISIS # 0 nM 10 nM 20 nM SEQ ID NO 129688 100 105 103 442 175510100 90 85 47 222035 100 88 80 244 298697 100 88 61 141

Example 36 Effect of Antisense Inhibitors of HIFs on Human Tumor CellXenografts in Mice

[0294] Nude mice are injected in the flank with approximately 10⁶ U87-MGhuman glioblastoma cells. Mice are dosed with antisense compoundbeginning the day after tumor inoculation and continuing every otherday. Oligonucleotides used are ISIS 129688 (unrelated control), ISIS175510 (HIF1α), ISIS 222035 (HIF2α) and ISIS 298697 (HIF1α with somecrossreactivity to HIF2α). Tumor volume is measured every few daysbeginning 10 days after inoculation.

[0295] Similar xenograft studies are performed with MDA-MB231 humanbreast cancer cells, which are p53-deficient. Nude mice are injected inthe flank with approximately 10⁶ MDA-MB231 human breast cancer cells.Mice are dosed with antisense compound beginning the day after tumorinoculation and continuing every other day. Oligonucleotides used areISIS 129688 (unrelated control), ISIS 175510 (HIF1α), ISIS 222035(HIF2α) and ISIS 298697 (HIF1α with some crossreactivity to HIF2α).Tumor volume is measured every few days beginning 10 days afterinoculation.

Example 37 Effect of Antisense Inhibitors of HIFs on AngiogenicConditions in the Eye

[0296] It is believed that antisense inhibitors of HIF2α and possiblyHIF1α will be useful in treatment of angiogenic conditions, because oftheir effect on endothelial tube formation in an in vitro model forangiogenesis (see previous examples).

[0297] A pig model of ocular neovascularization, the branch retinal veinocclusion (BVO) model, is used to study ocular neovascularization. Malefarm pigs (8-10 kg) are subjected to branch retinal vein occlusions(BVO) by laser treatment in both eyes. The extent of BVO is determinedby indirect opthalmoscopy after a 2 week period. Intravitreousinjections (10 •M) of ISIS 129688 (unrelated control), ISIS 175510(HIF1α), ISIS 222035 (HIF2α) or ISIS 298697 (HIF1α with somecrossreactivity to HIF2α) are started on the day of BVO induction andare repeated at weeks 2, 6, and 10 after BVO (Right eye—vehicle, Lefteye—antisense molecule). Stereo fundus photography and fluoresceinangiography are performed at baseline BVO and at weeks 1, 6 and 12following intravitreous injections to measure the neovascular response.In addition capillary gel electrophoresis analysis of the eye sectionscontaining sclera, choroid, and the retina are performed to determineantisense concentrations, and gross and microscopic evaluations areperformed to determine eye histopathology.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 458 <210> SEQ ID NO 1<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(3) <223>OTHER INFORMATION: 2′-O-methoxyethyl with phosphorothioate backbone<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (13)..(20)<223> OTHER INFORMATION: 2′-O-methoxyethyl with phosphorothioatebackbone <400> SEQUENCE: 1 tccgtcatcg ctcctcaggg 20 <210> SEQ ID NO 2<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(5) <223>OTHER INFORMATION: 2′-O-methoxyethyl with phosphorothioate backbone<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (16)..(20)<223> OTHER INFORMATION: 2′-O-methoxyethyl with phosphorothioatebackbone <400> SEQUENCE: 2 gtgcgcgcga gcccgaaatc 20 <210> SEQ ID NO 3<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(5) <223>OTHER INFORMATION: 2′-O-methoxyethyl with phosphorioate backbone <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (16)..(20) <223>OTHER INFORMATION: 2′-O-methoxyethyl with phosphorioate backbone <400>SEQUENCE: 3 atgcattctg cccccaagga 20 <210> SEQ ID NO 4 <211> LENGTH:3933 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: (265)..(2745) <223> OTHERINFORMATION: CDS <300> PUBLICATION INFORMATION: <301> AUTHORS: Hogeneschet al. <302> TITLE: Characterization of a subset of thebasic-helix-loop-helix- PAS superfamily that interacts with componentsof the dioxin signaling pathway <303> JOURNAL: J. Biol. Chem <304>VOLUME: 272 <305> ISSUE: 13 <306> PAGES: 8581-8593 <307> DATE: 1997<308> DATABASE ACCESSION NUMBER: U29165.1 <309> DATABASE ENTRY DATE:1997-04-11 <313> RELEVANT RESIDUES: (1)..(3933) <400> SEQUENCE: 4cacgaggcag cactctcttc gtcgcttcgg ccagtgtgtc gggctgggcc ctgacaagcc 60acctgaggag aggctcggag ccgggcccgg accccggcga ttgccgcccg cttctctcta 120gtctcacgag gggtttcccg cctcgcaccc ccacctctgg acttgccttt ccttctcttc 180tccgcgtgtg gagggagcca gcgcttaggc cggagcgagc ctgggggccg cccgccgtga 240agacatcgcg gggaccgatt caccatggag ggcgccggcg gcgcgaacga caagaaaaag 300ataagttctg aacgtcgaaa agaaaagtct cgagatgcag ccagatctcg gcgaagtaaa 360gaatctgaag ttttttatga gcttgctcat cagttgccac ttccacataa tgtgagttcg 420catcttgata aggcctctgt gatgaggctt accatcagct atttgcgtgt gaggaaactt 480ctggatgctg gtgatttgga tattgaagat gacatgaaag cacagatgaa ttgcttttat 540ttgaaagcct tggatggttt tgttatggtt ctcacagatg atggtgacat gatttacatt 600tctgataatg tgaacaaata catgggatta actcagtttg aactaactgg acacagtgtg 660tttgatttta ctcatccatg tgaccatgag gaaatgagag aaatgcttac acacagaaat 720ggccttgtga aaaagggtaa agaacaaaac acacagcgaa gcttttttct cagaatgaag 780tgtaccctaa ctagccgagg aagaactatg aacataaagt ctgcaacatg gaaggtattg 840cactgcacag gccacattca cgtatatgat accaacagta accaacctca gtgtgggtat 900aagaaaccac ctatgacctg cttggtgctg atttgtgaac ccattcctca cccatcaaat 960attgaaattc ctttagatag caagactttc ctcagtcgac acagcctgga tatgaaattt 1020tcttattgtg atgaaagaat taccgaattg atgggatatg agccagaaga acttttaggc 1080cgctcaattt atgaatatta tcatgctttg gactctgatc atctgaccaa aactcatcat 1140gatatgttta ctaaaggaca agtcaccaca ggacagtaca ggatgcttgc caaaagaggt 1200ggatatgtct gggttgaaac tcaagcaact gtcatatata acaccaagaa ttctcaacca 1260cagtgcattg tatgtgtgaa ttacgttgtg agtggtatta ttcagcacga cttgattttc 1320tcccttcaac aaacagaatg tgtccttaaa ccggttgaat cttcagatat gaaaatgact 1380cagctattca ccaaagttga atcagaagat acaagtagcc tctttgacaa acttaagaag 1440gaacctgatg ctttaacttt gctggcccca gccgctggag acacaatcat atctttagat 1500tttggcagca acgacacaga aactgatgac cagcaacttg aggaagtacc attatataat 1560gatgtaatgc tcccctcacc caacgaaaaa ttacagaata taaatttggc aatgtctcca 1620ttacccaccg ctgaaacgcc aaagccactt cgaagtagtg ctgaccctgc actcaatcaa 1680gaagttgcat taaaattaga accaaatcca gagtcactgg aactttcttt taccatgccc 1740cagattcagg atcagacacc tagtccttcc gatggaagca ctagacaaag ttcacctgag 1800cctaatagtc ccagtgaata ttgtttttat gtggatagtg atatggtcaa tgaattcaag 1860ttggaattgg tagaaaaact ttttgctgaa gacacagaag caaagaaccc attttctact 1920caggacacag atttagactt ggagatgtta gctccctata tcccaatgga tgatgacttc 1980cagttacgtt ccttcgatca gttgtcacca ttagaaagca gttccgcaag ccctgaaagc 2040gcaagtcctc aaagcacagt tacagtattc cagcagactc aaatacaaga acctactgct 2100aatgccacca ctaccactgc caccactgat gaattaaaaa cagtgacaaa agaccgtatg 2160gaagacatta aaatattgat tgcatctcca tctcctaccc acatacataa agaaactact 2220agtgccacat catcaccata tagagatact caaagtcgga cagcctcacc aaacagagca 2280ggaaaaggag tcatagaaca gacagaaaaa tctcatccaa gaagccctaa cgtgttatct 2340gtcgctttga gtcaaagaac tacagttcct gaggaagaac taaatccaaa gatactagct 2400ttgcagaatg ctcagagaaa gcgaaaaatg gaacatgatg gttcactttt tcaagcagta 2460ggaattggaa cattattaca gcagccagac gatcatgcag ctactacatc actttcttgg 2520aaacgtgtaa aaggatgcaa atctagtgaa cagaatggaa tggagcaaaa gacaattatt 2580ttaataccct ctgatttagc atgtagactg ctggggcaat caatggatga aagtggatta 2640ccacagctga ccagttatga ttgtgaagtt aatgctccta tacaaggcag cagaaaccta 2700ctgcagggtg aagaattact cagagctttg gatcaagtta actgagcttt ttcttaattt 2760cattcctttt tttggacact ggtggctcac tacctaaagc agtctattta tattttctac 2820atctaatttt agaagcctgg ctacaatact gcacaaactt ggttagttca atttttgatc 2880ccctttctac ttaatttaca ttaatgctct tttttagtat gttctttaat gctggatcac 2940agacagctca ttttctcagt tttttggtat ttaaaccatt gcattgcagt agcatcattt 3000taaaaaatgc acctttttat ttatttattt ttggctaggg agtttatccc tttttcgaat 3060tatttttaag aagatgccaa tataattttt gtaagaaggc agtaaccttt catcatgatc 3120ataggcagtt gaaaaatttt tacacctttt ttttcacatt ttacataaat aataatgctt 3180tgccagcagt acgtggtagc cacaattgca caatatattt tcttaaaaaa taccagcagt 3240tactcatgga atatattctg cgtttataaa actagttttt aagaagaaat tttttttggc 3300ctatgaaatt gttaaacctg gaacatgaca ttgttaatca tataataatg attcttaaat 3360gctgtatggt ttattattta aatgggtaaa gccatttaca taatatagaa agatatgcat 3420atatctagaa ggtatgtggc atttatttgg ataaaattct caattcagag aaatcatctg 3480atgtttctat agtcactttg ccagctcaaa agaaaacaat accctatgta gttgtggaag 3540tttatgctaa tattgtgtaa ctgatattaa acctaaatgt tctgcctacc ctgttggtat 3600aaagatattt tgagcagact gtaaacaaga aaaaaaaaat catgcattct tagcaaaatt 3660gcctagtatg ttaatttgct caaaatacaa tgtttgattt tatgcacttt gtcgctatta 3720acatcctttt tttcatgtag atttcaataa ttgagtaatt ttagaagcat tattttagga 3780atatatagtt gtcacagtaa atatcttgtt ttttctatgt acattgtaca aatttttcat 3840tccttttgct ctttgtggtt ggatctaaca ctaactgtat tgttttgtta catcaaataa 3900acatcttctg tggaaaaaaa aaaaaaaaaa aaa 3933 <210> SEQ ID NO 5 <211>LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR Primer <400> SEQUENCE: 5ccagttacgt tccttcgatc agt 23 <210> SEQ ID NO 6 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR Primer <400> SEQUENCE: 6 tttgaggact tgcgctttca 20 <210>SEQ ID NO 7 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: PCR Primer <400>SEQUENCE: 7 tcaccattag aaagcagttc cgcaagcc 28 <210> SEQ ID NO 8 <211>LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: PCR Primer <400> SEQUENCE: 8gaaggtgaag gtcggagtc 19 <210> SEQ ID NO 9 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: PCR Primer <400> SEQUENCE: 9 gaagatggtg atgggatttc 20 <210>SEQ ID NO 10 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: PCR Primer <400>SEQUENCE: 10 caagcttccc gttctcagcc 20 <210> SEQ ID NO 11 <211> LENGTH:57500 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: (1)..(57500) <223> OTHERINFORMATION: positions 82000 to 139500 of the sequence with GenBankAccession No. AL137129.4 <300> PUBLICATION INFORMATION: <308> DATABASEACCESSION NUMBER: AL137129.4 <309> DATABASE ENTRY DATE: 2001-04-30 <313>RELEVANT RESIDUES: (1)..(57500) <400> SEQUENCE: 11 taaaatttta tcctatatgaaattttcctt tttggtgtct gttatttaat aggattgttt 60 gaattagggg atactatttggtgcctttgt aactatatga aaattagttg gttgaatatt 120 actgctttcc atgttcatatttatatttgt atagacatat atatatatac acatatacta 180 ctttcctttc cattttcatatttatatttg tgtatacaca tatacataaa catatatttt 240 atacattttt gaaaaggaaaattaacttaa gggcatattt aatgaatatt caaaaatttt 300 tttgctgatc aaattatcattctgctttaa acttttgaaa tgatccaaaa aaattttaaa 360 tgacttagat ttactgttacaaaatgcttg tcttttgatg tcacaaacat tatatactat 420 aatcactggc cagagataattgctataagt ataatgaaaa gggaaatgat ggaagatctc 480 tgcagctatc ctcataaatgagggtgggaa cacgatgggc agttccaaag ttgaaaatag 540 agaatatatg tggatttatattaacataat tggtattctt ggatagttaa aaatggctaa 600 actgtaggag aagcccgagtaattactgtt aacagaggaa taaatttgag ggcaataata 660 atgatgatag gccaggcactgtggctcatg cctgtaatcc cagcactttg ggaacccgag 720 gcgagcggac cacctgaggtcaggagttcg agagcagcct ggccaacatg gtgaaacctc 780 gtctctacta aaaatagaaaaattatccga gtgtggtggt gcgtgcctgt aatcccagct 840 acttgggagg ctgaggcaggagaatcactt gtacctggga ggcggagttg cagtgagccg 900 aaatcgcgcc actgcgctccagcctgtggg ccagagcgag actccgcctc agaataataa 960 taatgataat aataataacgccaccaacaa tactaagagc taacatttac tgagtgctta 1020 ctatgcacca gatattgttctaagtataca tttattatct catttaacca tccataatac 1080 tgtggtatag acacttttatatccatttta taaataagta aactgagtta tggagagatt 1140 aaacgacttg ccagtaagattcaaagcctg tgtacaagct cacgcttgat tctggagcca 1200 gtgttcttaa cacagtatcttgagaatgtt aaactaaaaa gtttttaatt tacagtattc 1260 tttccacaat taaaaaagaaattatgagta attattttta gttctttctt ctcttcaggc 1320 atttcccatg gttcttttcaagacataata catatcattt agtgttgtag atctgaaaaa 1380 acaaaagtag cgtgaagatcaaaaattttc taaagagacg gagtctcgct acgttcccta 1440 ggctggaaca cccaggcttctccagcctca cacctctgag tagctggaac caccctgtcc 1500 gctaaggtca atgtttaatcgtatctttgt aggtctactg accagttaaa aagaggtgct 1560 gtatacattg gttgttgtcttgtcagagtt tgatgcttct atatagacca ttgtttttac 1620 atgctaatac aattgaaagccactacagat atttatattt acaacccaaa gctaggtttt 1680 aacaagaaac tcataaggcaaaggtgagaa gtaaaataat ttagcgccaa gtggagatat 1740 atgtgcaatg ctactttgttgggctcaaaa catatttttc ttttagaaga ctgacaggct 1800 tgaagtttat gcctccaaagacaaaagtga ttatgttttg tttagtagct tgcaaagttg 1860 ccaaaggcca ttttttctactctttccctg aaattggttt atatgcttat taaagtcatt 1920 tatacctatt tgcaaatgcttaacatagtt tcagatttta agatttccct gcaactttat 1980 ttcccttgaa gtttacagcaacaggagttc atttttattt ttaattgcat ttattcagta 2040 agtaaactcc gccacagaaaaacttagtag acaaggtgag ttcccctgtg ctccgtggca 2100 aagagtgcgg tgggtgacattgacccatgg ttaggtaatc tggtaaggaa agaccccgtt 2160 gtaacacatc tgagcaacgagaccaaagga agggcttgct gccacgaggc gaagtctgct 2220 tttttgaaca gagagcccagcagagttggg cggcaatcgt gcccagcact gaggccgagg 2280 agaaagagag caggagcattacattactgc accaagagta ggaaaatatg atgcatgttt 2340 gggaccaggc aaccgaaatcccttctcagc agcgcctccc aaagccgggc accgccttcc 2400 ttcggagaag gcgcagagtccccagactcg ggctgagccg cacccccatc tcctttctct 2460 ttcctccgcc gctaaacacagacgagcacg tgagcgtcgc agcccgtccc agctgtgcct 2520 cagctgaccg cctcctgattggctgagagc ggcgtgggct ggggtgggga cttgccgcct 2580 gcgtcgctcg ccattggatctcgaggaacc cgcctccacc tcaggtgagg cgggcttgcg 2640 ggagcgcgcg ccggcctgggcaggcgagcg ggcgcgctcc cgccccctct cccctccccg 2700 cgcgcccgag cgcgcctccgcccttgcccg ccccctgacg ctgcctcagc tcctcagtgc 2760 acagtgctgc ctcgtctgaggggacaggag gatcaccctc ttcgtcgctt cggccagtgt 2820 gtcgggctgg gccctgacaagccacctgag gagaggctcg gagccgggcc cggaccccgg 2880 cgattgccgc ccgcttctctctagtctcac gaggggtttc ccgcctcgca cccccacctc 2940 tggacttgcc tttccttctcttctccgcgt gtggagggag ccagcgctta ggccggagcg 3000 agcctggggg ccgcccgccgtgaagacatc gcggggaccg attcaccatg gagggcgccg 3060 gcggcgcgaa cgacaagaaaaagtaagccc attccctcgg cccgccgcct tctcccccgg 3120 cgaccccgcc cgcctgcccgccctgggctc ctgggccggc ctcggcgtta atgggattgg 3180 ggggggcagc ctttttgtttctgctgctgc tcccctcccc tctcttcccc caacctcgcc 3240 ggccgggctc ccccgctgtccacgtcgcca tcttgtcgtg gggggtggga gacgcctcga 3300 aagtgctttc aggggccggggtctgagccc tgcttgccct ccccgccggc cgtgggggcc 3360 tcgcgccgcc cacctacccgcctcaaaaac ccagcctgct ctgtggcccc atccggaggg 3420 gactttaccc agcctgaaaaccccgggaag agaaatgagc tgcagctcgg tagccgcggt 3480 ttgcacccgg agcttccgctccttcccgcc cccatcctct ccagttccat tgaaaactcg 3540 gccctggggc ggaccctgcacgctggtcct ggctttccag tggacttggg gccttgagtt 3600 cccgactgag ggactcgcgtggtcggatgc gatcttgtcc tgtagttgtc cagccgtcgc 3660 gggtgtcttt gcctttgtgcattagggatt tgccgcgatg gccttaagat gcgaactttt 3720 tagtttgcac gtgcaggttttgtttcgttt taatcgcctt gaaaaacttg cctagactga 3780 gagtcagagt aatgggaatttagggaaatg gcaacatttt aaagagaact tcagaattgg 3840 atacttgagt tcatatcacctgtcacgaga acgcagatat tataaatgaa tatatgcctc 3900 attcattctt caaataatgaaaatgtaggg gctggttaaa tttaggcagt tttaatgata 3960 ctgaaaaaag tatatgatgagtgaatgaaa tgcggcacta aaatgttgca aaaattttcg 4020 aactctgtct cattttcctgaaattgaagt atattaaagg aaaaccgtca acatatatct 4080 aaagtaagta atcactcggttagaacttaa tgcaagtttt ataaatcacc ttgaagtttg 4140 agtctaaggg gtacattagagattaagaat tgtgagttgg accagtggtg ttaagagcgg 4200 actcccccat cccccaacacacacacaatt ttgcccactt tggcatttta acttttaagg 4260 aaatcactta aggaattgaagatttagagt aagagttttg gttagtagac tggctttgct 4320 gttaaatcct tccactcttctggcagagag attaatttcc ctaatcagta tcagcagaag 4380 ataaacttgt ttatattcctgctgttttgt agatcccttc tcctggtcct tcttcaatag 4440 aatattaaat tcttagtttgtatacagcag agaaggtcac ttataaaatt caaaaagtga 4500 gcaaacaggt ctagattaattccaagagtt accaggaatt aattgcagtt tattttgcgg 4560 aggtgattac agtgcttttgatgaaatgat aaagctgcta tattgtaaac ctaaggcaga 4620 ttacctctgt gtagtgccagttttctatcc ttattatata ttgaatcata cttaatacaa 4680 tgcattaaat tatgtaccactttttttata tacagtatcg aactcattgt tttgccattc 4740 atccgttcag aatatcagaagcagttttga aacgaattaa taaattagct actgttcatc 4800 agccccaatt ctaaataagctcttagattt tcctcagccc atctgttact ttcaaaattt 4860 tctcatttga aaacttggcaaccttggatt ggatggattc atatttctta gtatagaagt 4920 tcttgatata actgaaaaattaagttaaac acttaataag tggtggttac tcagcacttt 4980 tagatgctgt ttataatagatgaccttttc taactaattt acagtttttt gaaagataac 5040 tgagaggttg agggacggagattttcttca agcaattttt tttttcattt taaatgagct 5100 cccaatgtcg gagtttggaaaacaaatttg tctttttaaa agaaggtcta ggaaactcaa 5160 aacctgaaga attggaagaaatcagaatag aaaatgggta tggttatgat actgtagatt 5220 taacgcagga catttcatgttgttcctagt tataggggct gaacttattt aatagcacgt 5280 gcattttgat ttttagatttttaagggaat gtcaagagag taatgattct gtttcaggct 5340 tcaggccaga ctccttcagagttttccaaa acaaataatt actgaatcat taaagtaaaa 5400 tttctgagaa tagatattccttaatttcct tcattaactt tggccattaa aagtcaagaa 5460 gctctctcat ttattagcaaacttttctcc ttatgattct attttgattg tccttttgtt 5520 tgaggaagca gcatatggtggttaagagca taggatctag aggcagatac ctctgagtta 5580 agggtcccag cccttcacttgtgagcttga gcaagttact gaatgcctct gagcctcttt 5640 cctccttttg aaatgatgataagaatagca gccatctgag cagttattgt aaaggttaaa 5700 tgagataatg cttgtgaagcacttagccca ttgcaggagt cttgatgaca ctgtgtactt 5760 gaaaatagat gttacctgttaaaattcttg tttaaacttc cacaactctt aaaactcttt 5820 tttgctagtc cttccagctgtttcctttag tttcttttct gtgtcttcat gcatcttttc 5880 tatctcctga aagtgaaaagactaacattg gatccagagc ttgaaaagcg tttttttcct 5940 gttacaatgg gcaaaagagtacatccttgg gttatattgg cacctagtat cagttatttt 6000 tcttgagcat ctgatctgctctctactcta gtggaggcct cctgcttcac aattgctcac 6060 ccctgtgttt tctccccaaatagaatactg agtttactct ggactctaga gtcaaacata 6120 cacagtattc tagtcttactgttcatttaa gcaagatatg tgcaagacac tgcattctta 6180 gtactggcag taagttaaaacatttttcgt cttgatgcca aagtttagac aattttataa 6240 aaattaacct ttgtaaaagataatgagttg ataaaatatt ctcagtaaag cagctacgtg 6300 gtagaaaaac tgtcctttgcttatgagttt ctccagagtt aagaccattg ggttccatct 6360 gaaggcaaga cttcaagcttgtcttactgg tctgttttgt ggctcaattt gtatgaagtc 6420 tatgcactct tccacacgtgtgtatttact gaactatcga gttattttag actgagaaag 6480 tattggagtt cattcctacggtccactgca gagcaccttg tgcagtttgg agaatgtcaa 6540 cttttctacc tgttaacttccattgtcttt acttttaacg ccattgtctg tgactctaat 6600 ggtgtcacgg ctcagggtttagattttgtg gttacattct attcttgtat gtcaagagtg 6660 gtgtatagaa agctgagggggattatttag tctcttgact gatttttttt ttttttctga 6720 agaactcagt ttattatgtttggtggtgaa ataaaaattg atgtgcatgg atgttaaaga 6780 tttgggttaa attgtgtgttcatagatgcc ttctcttagt atataatttt ttaaatttag 6840 atacttaaaa tactgtatccctttatctaa gattaacata agtctgtttc ttaaccagga 6900 taaaaaaatc taaatttaaatgtgatgttg gatgagtttc caatcaagaa attgattttt 6960 taaactttgt gactagttatccagtgggtg gattttaccc agtgtgtgta tgtgttttct 7020 gcttaactct ggaaggttagaaagagaatt tgaaactaag acaagccaag cttcttgttg 7080 ctcagtattt ttggtaaaaatatggtcaga ttgtttaaat taactatagg ctttggaatt 7140 ttaaaaataa ttatatctcttggtctcttg acacatcaag aattaactgt tttgtatatg 7200 cgttgagtat taatgttcatgttttctgca gtagaaattt ataaaccctt atttatttgc 7260 cagacatgat ccctttagagaaatctagta tctaaaacct gaatttttaa aacaaaattt 7320 aaaatttttg tttcataaaaacaaaaatgt gattacctca tggctttttt cttatagctt 7380 ttgattgttt tttaaaatcgtagttcaaaa acattaacct aaaatttacc atcttaacca 7440 tttctaagta ctgttcagtagtgttaagta tattcacatt gtgccactaa cttccagaac 7500 tttttcatct tgcaaagctgaaatcttacc cattaaacaa ctcccaattt ccccctctcc 7560 tcagcctctg gcaaccaccattttactttc tgtttctgca aatttaacta ctctagatgc 7620 ctcatataaa tagaattatagggttttaat atttttgtga tgggcttatt tcactttgtg 7680 taatgtcctc aaggttcatccatgttgtag catgtgtcag aatttccttc ctttttgagt 7740 ctgagcaata ttccattatatgttccatat tttgtttctc cattcatcca gcaacggaca 7800 cttgggttgc ttccacatcttggttattgt gctgctctga acatgagtct gcaaatctct 7860 ctttgaagct ttcactttttttggatacat atccagaaga gggatgctgg atcatatggt 7920 aacccttttt aattttcaaggaaccaccat attgttttct atagcagttg caccagttta 7980 cattcccacc aacagtgcacaagggttcct atttctccac atccttctaa acacttgttt 8040 tctttctttc cttccttctcttctctttct ttcttagcca tctaatgtgg caaagtggta 8100 gccatctaat atgttgaagtgattgttttt aagggcttgt ttgtggataa ttaaccagct 8160 gaaagctaac tacagtttgccagtggaagc tttaactgaa aggagagtaa gtacctctaa 8220 aaggagaatt caatttttctagtgacttag atttgttatg ccagtacttt ttcacagaaa 8280 cactttttgg gtaaaatagtgtacacctgt tctattgttg ataaagccca atttaattag 8340 gaaatttgtt ctctaagatttaaaacaata attgaaataa tgtattttta ttaaaaactg 8400 ttcccaagat gttagcttttagctgttctg gtgatctcaa ctgttattta tgagtgtttc 8460 tttattttaa aatttcaccttaaccggtta cagttttaac cataaagatt atttcaacat 8520 atgattttga aaatttattatcttgtaaat gggaaaatgt agtgatggaa catagtttac 8580 tgtatgtagt tcttcacttgtttgaaaagt cacaatatat ttaggcaaat taatttaaaa 8640 gtgtctagta tttaatattgcaattttcac tcattaagga caggtccccc gtgtttcccc 8700 cttttttttt tccaagtagtttgggaggat ttgtttttcc agctgaaaaa tactatggtt 8760 aaaaataagg tttaaaggcgaaagttgaag tctttgaggg ttgggatacg tttctgttct 8820 taagagtctt gtaaattcagatgctaagca aatttcttta aaatgatttc taccctcccc 8880 ctttccatta taaaactggatatgtttcag tggaccaaat cccaagtagg ctgaatttga 8940 aatttgtggg ctgggcgcggtggctcatgc ttgtaatccc agtactttgg gatgccgagg 9000 tgggtggatc acctgaggtcaggagttcga gaccagcctg gccaacatgg tgaaacccca 9060 tctctactaa aaataccaaaattagccagg cgtggtggcg ggtgcctgta atcccagcta 9120 cttaggaggc tgaggcaggagaattgcttg aacctgggag gcggaggttg cggtgagcca 9180 agatcgcccc attgcactccagcctgggtg acagagcaag actgtgtttc aaaaaaatta 9240 aaaaagaaat ctgtggtgtgaatactggta cgtggtgtac acagtgagct cttaataagt 9300 atttgaatta acaaatgagacaatgattga ataattggat gaacaaagag aatgcaggtt 9360 tttaaaaggt ttctttagaaatattgtcgg cccggcacgg tggctcctgc ctgtaatccc 9420 accattttgg gaggccggggcaggtgaatc acctgaggtc aggagttcaa gacaagcctg 9480 accaacttgg agaaaccccgtctctactaa aaatacaaaa aaaaaaaaaa aaaaatagca 9540 ggatgtggtg gcacatgcctgtaatcccag ctactcggag gctgaggcag gagaatcgct 9600 tgaacctggg aagcagaggttgcagtgagc caagatcgcg ccactgcact ccagcctgat 9660 gacagtgtga gatgctgtctccaaaaaaaa aaaaaaaaaa ttaaaaagaa tgttttaatt 9720 ctttagttcc ctgtctgagattcactgatt ggtaagaaga aagttaaaga atctcctttg 9780 actttttttg atatagatatttaaattcta ttactttata gtaaggttgg ggtttatttt 9840 ctttgcttta taatagaagagcattgatta ttctctttgc tttataatag aataccattt 9900 aaataggagt tccctgagtgtgtttacaat catttgatct ggctaaacta ttttaatgtt 9960 aatgaaattt taaaattttggaggaaaaaa tttaaaaact acacaggtgc acaaagaaat 10020 aaaaatcacc tgctttttcactatgtagag accattgtct actatttctc aattctgtgt 10080 tacatctgta tgttaataactgtaggatta gggactgagt actgttttta acctgctttt 10140 aaaaaattta catctacattttttcccatc taaatagtga ggaagagtat cagaattttg 10200 taggcttgtg gtgatggttaaattagataa tattaatgtt gggtacttaa cataatatat 10260 ggctcttaat actctccagatttcagatat agtctgtttt accattactg cctttttatc 10320 aaacctattc tcaaaaaagtgagaaaagtg ctgagattac aggcgtgagc caccatgccc 10380 ggcctcatgg ttctttcttaataataaatt agaagaagta gaattacagg gtcaaaaagt 10440 atccatttta aagctttcaatgtaattgcc tgtttatctt ctagaaagtt tgacctagtt 10500 gtattttaga gtgtcattttcttgaacttt atcatcatta aagttttaaa tttggaacac 10560 tggcaatttg ataagtatattaggattctt cttattgcaa gtagcaaaat acaactcaat 10620 ctagtttaag aggggaaaatgtagtcattg gctaacacaa tctaattttg gtttaagaga 10680 caaatctaga gtctcaaatgatctcagagt gtaataatcc ctgacttttg tcttgatatt 10740 acttggcttg tatacctttgctctatttgc atgctggcct tactctgcca ctgacaggct 10800 gtctgtatgg tgtggaagaggacggctagc atccccatac ctgcatccat acagtttgta 10860 atataaaaaa aaaaaaagtaaaaaaaactc cctctctctt ctagtgtcta tatatcagtt 10920 tcctagaaga aaacgttttgccctacttgg ccatgtgaat ggagttccct gattacatga 10980 gtcaaatatg tcttattgtagcatatttga tggtcttctt gtagaatatt atcttactat 11040 acacagaact cttgaccagtaattaatggg ccatgagttt ttgttgcaag tcatttgaat 11100 tcatattcta tagttttctaccaagtgtag tcattctgca agctgttctt gtcatgactt 11160 ttgggaagtt gagtatttcttctatgggtt agggttttca tctcaagaaa aagatgatcc 11220 ttttctctac taaatatgtgttaagatcac acatttttct agatcgttta gctctactgt 11280 gtgatcttac acaaattgctttattgggat gataagaata attgccttat aggattgtta 11340 tgagaatgaa atgatacatcaactcatatg aaacactcag aacagctctt ggcacaaagt 11400 aagggcttaa ttaagtagaaactatccata tattcataat attatagtat tggttaagtt 11460 gttttcaaca ttgtttagaatcgctcaagc cttctttgtg ataatctgac gaaggctatt 11520 caccaccagt gagtaaataatagtggcaga atagttactg atgcttttcc tttacttggt 11580 tttttttcca taaacatctggcctttgcag actaaatact ggtttatgta tagacatgtt 11640 attctaaaat aattttccatagtggtaata ctaaaggaag aaaaatgttc tcaaagctat 11700 ttatttggga tgttaaaggagggggaaatt aagaaagcct acatttccat gtcctttgtg 11760 tccagaatct cattaaatgtcttttaactt gttagcagag gaaagttgga tattgcctgc 11820 ctttgtagct aacatagttaaaatatttaa atggttatag tgtcaaacca gtagtcaaag 11880 ccttcactgt gaatggatgaagggatattt tcttgaataa tttaagttga cttatttcag 11940 tggttcaaaa aatttcttcaacgcttaacc atgactcagg cacctaacta ttatactatg 12000 tcctgtaaca gattgttgtgcattcattta ttcaacaggt atttgtgcag ctaatttatt 12060 gagtacagca ttgaatcgttgatggcttag gccacagttg aacattccat tttttatgtt 12120 cattcattca ttcatagcatattccatttt taaattttca gttcattgca ctttaaagtt 12180 tgaggttctt gcgaagtacagacttttggg tttaagtttt gttatttaat gtcaaccacc 12240 acaggcgcat tggccagtctgcttttagaa ttttcagaca tacatacaca aaacattctc 12300 acaagacaat ctacttattttcttttttat tcctgtgttt cttaacacag gattaatgtt 12360 cagatctctt ttggagcaaaataatcctct gaatttttga gatgtaccca gtgacctcag 12420 tctgagtatg tatactgcattaaaaaatgt aaccttgttc cttttagtgg tcatttggta 12480 acagtttgat cataaacaaatgcagcctca aacacagaag gcttgaggca agtatacaga 12540 actatggaga gatcatttagatgatgtaga atatgccttt tcttttttta caatgccacc 12600 aaaatgaaaa cacggttttaaaaattctca tagagtgtaa cttcaacact gctttaactc 12660 tattaaacaa agcactgccatgttgtaatt cctatttatt actctctgga gttgtataaa 12720 ttaccaaatc cgccttttgtttgatatcct tttcaaatat ctgagggtag ctatcatgtt 12780 tcttccttct attcttaaaaaatagtccca aatttcttga atcttttaat ttaaaaatta 12840 tatattgagc atctgatttgtggaaaggca taggccatat taaaaatggg gcttcatatt 12900 aaaatgggga aaagggtggagattctcagg tggaatctga gatctgccac acactaatag 12960 tgttacctaa ccctttttaaagacaaagaa acaggatcag aaggtcactt tggaaaattt 13020 atttggtaat attggataggatggattagt atagttggaa aacagagact cttgctttag 13080 gagagctgct cctttgtcatttccagaatc ttaatcatgg tcaaggttta gagctaaata 13140 tttaatagaa gaagtctttagggtatgctt tctattgtac acccttattt caatacatgt 13200 gttttttcct gttatgtaagtactttatta ttatttatgc atcttctatt aaagttaagc 13260 aaataattat ttcaaggacacattcttcta catacacaca aagtttaggg tcactgacct 13320 tcttaggttc tagtcttagatctgttacca tctaagagca tataaataag ggaaacagaa 13380 agaaaaggat ttacaagctgagaaggaagc aatgcagaga aagaagagtg atagagtagg 13440 taatttgggg aaagtcagtgatacacagct cttaaccatg aacagtgatt cttcactctt 13500 gaatgtttgt gacattcatgaaggtattaa aagctgactt ttaaaaaatt gtttcagaga 13560 actggaaaaa aattcagttgccacattctt ccttaggtca tctttgaact ctactcatgc 13620 acttacgtgt ttaaggcaaagttttactaa acgcacactt gttcttgctg gcttattgac 13680 ttttactgct agcttcttattcttagcaat tatacctcac attacatagt attgtgaaac 13740 tcactatatt cagtgttttgcctgacaaac atggtatgtt ataggatgtg tattcagtta 13800 tagctaaaaa taaattattctcgtttttca aaatttgctg gcctacctgt taagcttttg 13860 ctttaagacc tgctaatgtttctcaaactt ctgtggttaa atcacctgag tgtctagttg 13920 ctctatggat tcccagggacccattcgcca gagattctga tttggtaatt ttgggatgga 13980 actcagggat ctgtaaattttacaagcact cagaaatgaa acatagactt taaacagcta 14040 agagtgctca tcaggattatgttgatatta ttttttaaac agatgtgcca agcctttaat 14100 ttgaatttcc agggttgggatttggccttc tatatttggg ggaaaaaagt tctattgatg 14160 attgtggata tataccacaggtcaaccatt gaatagtcta gtcagtgtag ttagtgtatt 14220 ttataattac taagttctaagtatgtggtg tattaatgtc ttaggaggtg gatatatttc 14280 ctgtatttgt aaagcatttgggtaggtttt ttaaagagaa aagtatgtaa caaactagtt 14340 ttgagcgttg ctcttttacttctttgggca tttttgaaga acacgttaag tatcttctta 14400 gagcagaggg gctcagagtggtccccagat tatcatcatt ggtaacacct agttggtgca 14460 ttactaactt gttagaaatgcacattctca ggcgccattc agacttcata aatcagaaac 14520 tctggaagta aggctcagcattctgtgttt tttttttctt tattatactt taagttttag 14580 ggtacatgtg cacaacgtgcaggttagtta catatgtata catgtgccat gttggtgtgc 14640 tgcacccagt aactcgtcatttaacattag gtatatctcc taatgctatc cctccccgct 14700 ccccccaccc cacaacaggccccggcgtgt gatgttcccc ttcctgtgtc catgtgttct 14760 cattgttcag ttcctacctatgagtgagaa cacgcggtgt ttggtttttt gtccttgcga 14820 tagtttgctg agaatgatggtttccggctt catccatgtc cctacaaagg acatgaactc 14880 atcctttttt atggctgaatagtattccat ggtgtatatg tgccacattt tcttaatcca 14940 gtctatcatt attggacatttgggttggtt ccaagtcttt gctattgtga atagtgccac 15000 aataaacata cgtgtgcatgtgtctttata gcagcatgat ttataatcct ttgggtatat 15060 acccagtaat gggatggctgggtcaaatgg tatttctagt tctagatccc tgaggaatcg 15120 ccacactgac ttccacaatggttgaactag tttacagtcc cactaacagt gtaaaagtgt 15180 tcctgtttct ccacatcctctccagcacct gttgtttcct gactttttaa tgatcgccat 15240 tctaactggt gtgagatggtatctcattgt ggttttgatt tgcatttctc tgatggccag 15300 tgatgatgag cattttttcatgtgtctttt ggcagcataa atgtcgtctt ttgagaagtg 15360 tctgttcata tcgtttgcccactttttgat ggggttgttt ttttcttgta aatttgtttg 15420 agttcattgt agattctggatactagccct ttgtcagatg agtagattgc aaaaattttc 15480 tcccattctg taggttgcctgttcactctg atggtagttt cttttgctgt gcagaagctc 15540 tttagtttaa ttagatcctatttgtcaatt ttggcttctg ttgccatggc ttttggtgtt 15600 ttaaacatga agtccttgcccatgcctatg tcctgaatgg tattgcctag gttttattct 15660 acggttttta tggttttaggtctaacattt aagtctttaa tccatcttga attaatttta 15720 gcataaggtg taaggaagggatccagtttc agctttctgc atatggctag ccagttttcc 15780 cagcaccatt tattaaatagggaatccttt ccccatttct tgtttttgtc aggtttgtca 15840 aagatcagat ggttgtagataagcggcatt atttctgagg gctctgttct gttccattgg 15900 tctatatctc tgttttggtaccagtaccat gctgttttgg ttactgcatc cttgtagtat 15960 agtttgaagt caggtagtgtgatgcctcca gctttgttct tttggcttag gattgacttg 16020 gcaagcattc tgtgttttgagaattcttcc aggggactgt gatgaaaact gacgtttgag 16080 aaccttcatc ttagagtaaaaactttacat acacattttt gttgttttat ttatctagca 16140 caatacttct tttttttgaaatggagtttt gctcttgttg cccaggctgg agtgcaatgg 16200 tgtaatctca gctcaccacaacctccatct cccaggttca gttgattctc ctgcctcagc 16260 ctcccgagta gctggggttacaggcacgtg gcaacatgcc tagctaattt tgtattttta 16320 gtagagacgg ggtttctccatgttggtcag gctggtctcg aactcccgac ctcaggtgat 16380 ccgcccacct cagcctcccaaagtgctggg attacaggcg tgagccactg cacctggcac 16440 aataccttat atataatcagggctcaaaga tttgttgaga ggctcaacac caattctgga 16500 ccaggaaaga ttttatttatatcactagtc aggaataatc taaaaacaaa aagcacattc 16560 ttcttacaag taatatttcaatacacatta atgtaaacac atggaaaagt attagctact 16620 taataaatta acatgtaaatgaaaaattta cacattatgg ctatttcaga tgtgatatag 16680 atttcatttt cagaaggaaccctccaatgt aaaacagtga ttcttttccc cgtttatttt 16740 actgcattag aaaatcacatttaaagtaag cattttggtg aggtttggaa ggtgaataaa 16800 tccatctttt ctttaattatggatatttaa gagagatgtt gttgtgccgt ttagataata 16860 atgatctaaa ccaagaaatttagttgcttt caaaaataaa ataagtgtat gcattctgaa 16920 catttttctt tagaaacaaaccatttcatc tgtttttttg aatttcaaat taattataca 16980 gaattttcaa aatttgaaaattaggttagc atgagaaact gaagatactg aattatattg 17040 cctgttcagt ctatacttttctttaggata tacagtagga aagaaatatg atagttcaag 17100 ttagattact acttctttcagagttttttg acaaatgcag gtacagtgat agtgtcagtt 17160 catggtgaat ttttgttaaaataaattaca aaaaatttgt gatcctggta tcttgaaact 17220 agttaatatt tgtaaactttgctaacactg tatatcactg tattctggtt ttatctgtgc 17280 atctatgagt tatatgtgtgtatagctaca tatgtttata tttatacaca tacattacac 17340 acaggagtgg aatcatactcaatttttttt gtatagcctg ctctgttcat ataatactat 17400 attgtagcat ctagtataagcaaagattaa tttttgtaga ctttgctttt atcctgaaat 17460 tttgtggtag ctggtttaatggaaagacaa tttctgtgac gtgttttgtc agttagggat 17520 tgaccctggt aaaatattgctggataacaa caagcaatgt aaaaatacat ttgttccata 17580 agataacctc cgtgaaggtagagacttggt ctgttttgtt tattgcaccg tgtcctgttc 17640 tgggaagagt gttagactcatagaagatga tcaagaaata ttttttgaat acatcaataa 17700 cattctctaa catgtgggtatcctaaaggt ttatttttaa agtttattga ttagaattca 17760 gaagatattt tcccagataaaataatagat tgctagctgt cttgaaaatg taatttatat 17820 ttaatttgaa atgtcaggtttttgctattt tttccattaa gtagagatag ggtttttaaa 17880 aattacatgt gatgttttaagtattctggt tttgcaacaa ttactagata gaaaatgtaa 17940 caacagatcc tattaataatacttccaata atacatataa aatacttgtc taaaagtaac 18000 cctccttaaa aaaacaaagctggccaggcg cggtggctca cgcctgtaat cccagcactt 18060 tgggaggctg aggcaggcggatcaagaggt caggagttca agaccagcct ggccaacata 18120 gtgaaacccc atctctagtaaaaatacaaa aaattagccg ggtgtggtgg caggcgcctg 18180 taaccccagc tactcaggaaaatcgcttga acctgggagg cggaggttgc agtgagcgga 18240 gatcgcacca ctgtacttcagccttgggca acagtgcgag actctgtctc aaaaaaaaaa 18300 aaaaaaaaag gcaataggattaggtatcaa cttaatgaaa acttcgtgac agcactttct 18360 tgaaaaagac tgtggaaaccaaagttagta aactcctgtt tctgcctggg ttcggaaaac 18420 ataaagatga taaagatgtttaagtattcc tttttttttt tttttttttt tttgagacag 18480 tgtcttgctc tgtctggagtgcagtggcac aatcacagct cactgcagcc ttgaactcct 18540 gggctcaaat aatcctcctgcctcagcctc ctgagtatct ggaactacag gagtgcacca 18600 ttacactcgg ctagtaatttgattggttaa gaacattaac tataactcac acattttcct 18660 gaccacattt gcttaggacaaaacagtaaa agacatgagt gtagatgaaa gcgataaggg 18720 aactaatctt aaacactgaacctcttttca gcaaattggc tttctagttt ctcagctctc 18780 tctttacacc tctaaatctctttcctggca agatcattta tttgccttgg tttatggtga 18840 tactcttcat tgttatactggtgggtgatt gttttaattg atagctgttt ttttctactt 18900 caggaagatg acactgctggctctgctggc tctgatgttt accttgtggc taatgcctgt 18960 gtttgcctgt gttcacatttattccacgat tcatttgtta acatttacta agctgctttt 19020 ctgtgccagg aacttggctagataaataaa tggttgtttt tgtacacaga attagctgtc 19080 ataatcagtt actgtagcatttattcttgc aaaaatatat atttatactt caactagtga 19140 tcgaatctca acttattaattcatacattc agccagcaca taattgaata cttcttatgt 19200 gtcagaaact gttctaggtgcttgggatgt tcattgaaca aaatagacaa aagtctccgc 19260 ctctatggaa cttactttccagtgaaggtg tggattggtg ggatagaaaa taaaataatc 19320 aagtaagata tgtacttaggctttcataaa aatacagcag ggcaagagga ccaagatgga 19380 ggcagtgatc agggaatctcaatgagggtg agactgcgac aaagacttga aaaaggtgga 19440 gaagcaagcc ttgtgggtatttagggtagc agtagtccag gcaaggggaa caactagtgc 19500 aaaggctcta ggaggcaatgtgtttgaagt gttttaagaa cagtaaggag gctagtatgg 19560 ttagaacaga atgagcaaagggggcaaagt ggtagaaggt gagatcaaag aggtaatgag 19620 gccattgtgg aggcccatatggactattgg aagggctttg gcttttactc taaatgaggc 19680 aaaaaccatt ttaagcagagaggagtgata tgacttgatt tcttgttaaa aggattattc 19740 tagttgctgt tacagaaaaagattacaggg gtgcaaagaa acagggagac aaaagaatat 19800 aagattttca ctgtaacttatatctagtat gcttgcttat acttgaaaat gcatatccag 19860 ataattgtag taaattcaaatattatgttt atttaatagt actaacattg atatgctggt 19920 taattatgat taggagcactaataaagcac aaatcaggga ttcccaaaaa gaatgttgaa 19980 agggcagtca gcttttcctgtgccagaaat caaagtcata gcagatttgg ggcaaatatg 20040 tcaaagtcaa acttacgcacatcactactg agaagacaaa gatgaatgtg tgacagtttc 20100 ctgcccccaa gaatctttaagcattgtgaa ggaagattaa tatagccaaa taactagagt 20160 gatcagttct accagagaggaccagttttg gaagccagag gaaaaaaaaa aaaaacagaa 20220 acaaaatgat gtttgaattaaatctttaaa agtttctctt ataaatttac caagccacat 20280 attgggaatg gtaccccaggcagaaggagt agagtaagca agccagaaag gaaatactat 20340 ggtgcttttg agtaactgcagtgtggctga agaatgtgga aaatgatgag gataaagagg 20400 tggacaggga actaggtaagggagggcttc cttttaaata attagacctt gtcctgtgta 20460 catttaatgg gattttaatcaggccataat gccaaatttc tttacttcgg aaggatcttt 20520 atggtgatgg tttcagaaagaaattaaagc agagtaacag tggttagcaa taatgatcag 20580 ctagtggttc ccaaacttacgtatcatatg catcttggaa gtttttaaaa actcagattt 20640 tgggatcctg acttagatctactgaatcag aatttacaga ttcaaattcc cagtgaggcc 20700 taggaatttg aaatgttgaatgtccttcac gatgcagcta gacaagcatt tgggaataaa 20760 gcattaggtg actatttcagtagactaagg agtgggaggc catttaagct caaaggctat 20820 tctacttctc actatatttctagtacctag cacagtgcat ggtacttgat agatgcatcc 20880 tttctcccat acctcgccctacacatctct tcatgtgtat ccttattaat atcctctatt 20940 ataaactggt aaacatgtttccctgagttc tgtgagctgc tccagcaaag atgggtttgt 21000 gagaatccca acttttgaagcctgtcagtc agaagttcct gaggccagac ttgcaactcc 21060 tgttgagggg gcagtcttggggactgagcc ctcaacctga cactgtctcc aggtagatag 21120 tgttagaatt gaattgaaggacacccagtt ggtgtccgct gcagaactga ttgctcacct 21180 ggtggtggag agaacccctcctctcccgat agggttgcag aagttgtctt ctgtgttgtt 21240 gattgctgtg gtgtgggagcagagggggga aaaaagctgt tggagagttt tttccaaaac 21300 aataggagat tatttagatttataaaaata gaatcaaagt agattaactg agcacattgt 21360 gaaatataga gtagagctgtgtgtaaggag tatatcttaa tgtcaagctg acaccaaatt 21420 gaatgtttgc tggaacgttcaaaaatctaa gcttcccaaa tctgtgaaaa cactcaggtt 21480 agtaaacagt cttatgcaaacagcaagaca atgctcaaag ccatttaagg aaaaagaaca 21540 gtaactgaat tctcttatggaaatgtgaga tgttgtttta gtaagtactg atggtgttat 21600 actttttgtt tattcgtttgctggtatttc agttcctaaa attccttcaa atatgctgca 21660 aaatacaaac caagaacttggtggattttc catttgtttt cctgtgggaa atgatggaat 21720 taaaaacctt gaggattagaccttgagagt taccttccag tgtttatgcc accattatac 21780 aaaattctgg aggacaaaacccttcccact taaaaaccag ttagtttcag aaaatcacct 21840 catgttagga gactgcatcattatagtatg tgtgttagct ttaggtatag atctaaaata 21900 tttttaatat tttaaaaacttaagcctttc ttcattaatt tggcctaata caagttagaa 21960 taactttaaa aatgagtacaaacaacaagg aagggccagg cgcagtggct caacgcctgt 22020 aatccgaaca ctttgggaggccaaggtggg cagatcacct gaggtcagga gttccagacc 22080 agcctggcca acataatgaaaccccatctc tactaagaat acaaaaatta gctgggcgtg 22140 gtggcacacg cctgtaatcccagctactcg ggaggctgag gcaggagaat tgcttgaacc 22200 caggaggcag aggttgcagtgagccgagat cgcgccattg cactccagtc tgggcaacaa 22260 gggtgaaatg ccgtctcaggaaaaaaaaaa acagtttctg tgactgctag acaaatgttg 22320 agcaagtaaa acaccaacaatgttgaactt agatattgaa atagctgctc tgtacaaata 22380 aagtctactg ggagtatagactgaattacc atcttttgac tctttcgcca taatgattgg 22440 cattaccgga agggattaccttgctttgaa gagctgctgg acagtagagc agagagcatc 22500 tattaccatt gtaggtgcctttcagttagg attttggatt tataagcaaa ctccaagaaa 22560 gagcctggtt ctgagtttctctgaatagct taggtcaagt cctaaattct gaagccaact 22620 cctataattc cttctttatgtctttggcat gtgaagtagg caaatttcga actttataat 22680 aatagcctag acttacaaatacttgccttg gtaatcagga tgagtttttg agagacaaca 22740 tagtctagtg ttaatcgcgtggacaccaga ctgcttgagt gaaatacagg ttctaccatt 22800 tattaacgga gtaatgttgggtaagctatt tagccagggt ccttatctgt aacatggtga 22860 taataataaa gattaaataataggtgaaaa atgtttagaa taccactgtg ttattagtaa 22920 gcaccatgca taggtgtttggatttaaaaa tactggcaaa ggccaggttg ggtggctcac 22980 acctataatc ctcgcactttgggaggccaa ggcagaagga tcgctttagc ccaggagttc 23040 aggaccagtc gaggcaacatagattccgtc tctgcaaaaa atttaacaga attagttggg 23100 catggtagcg tgtgcctgtagctacttggg aggctgaggt agggaggggg aggattgctt 23160 gagcccacga tttcgaggctgcagtgagct tatgatcatg ccactgtact ccagcttggg 23220 tgacagagca agactctgtctctaaaataa aatgaaaata aaactgcagg caaaaatgcc 23280 aactgaagag tgaacatgaacttttctttg catttttctt gggcctgaga ctttaagaag 23340 tgcagggcag ttaaaatgatgagatataat tctcacctat cagctcagca gaaattaata 23400 agattaaaaa gatgcgtaatatataatatt gcagagtgca tgggggaatt gatatacaca 23460 ttcatgaact ggcagagacaaaaatgggca cagaaccatt tggaaagcta ttgtgtattt 23520 taaaaaattt cagtagcacattttttatat catgaaattt cacttcagaa tgtcagtcct 23580 gtagaaatac tgacgcaagtgcaaaaacaa caaaaaccaa cttgtacctt caaggccaga 23640 aagagttatt tcaccaaataacataattga ggtacattaa ctttattaga agtaaatctg 23700 ataatctgct cacattttaaatagttatgg tttaacttca gttcttgaag tcacatattt 23760 ttacaattag gaatgctaacaggctttttg tgcaatacga aaagatgact ttaaatgcct 23820 acaattattt tgtgtccttttatttttttt taatttttac tgacctacta caaagcacta 23880 aatattttat gttcttaatctgaagaacaa tagacattct ctataaaaca actcttgctt 23940 attcatgaac tttgtacacaagaagcttaa taagacgggc tcaaaattat ttttctaaat 24000 atatttccta tacaaaataatttcaagata taattgttac ttttgtgtct aatactgtat 24060 gttaaataat aaaatggtaagcatgtaaaa actacaatac cacaaagatt gagctatttt 24120 gccagtagta tactccaactttagttctag aacagttgta gaaatgggta aacaaactgt 24180 tttaactgta ctcttaactgaaatatagta ccttatgcag tagcagaaca tatcagcaga 24240 agaacttcac ttgacctgtacttaaaaaca aaacagatgc aatttataaa atttagagaa 24300 atatagtgac cttatttgcatgtggaaaat gtacttcttt ctgatctaca tatcttctgt 24360 tgtgcaatgt aagcagtaaaacaaatagta caggattcat ctctgtggga cctagacccc 24420 ctggtctaac aaataattcttggtcagtac tgtaattctg tggtataaaa ctgataaaat 24480 tagccttcct gtgactagacaagaagccgg gcagtttaaa tgctgaaact cacaagaact 24540 tcagaagctt tagctttaagctttaagctt acttagaaat gttataagac ctccagtagt 24600 cacatatgaa gaatatcatgaagatttttc cattaaatct ttattataga tcccttgatt 24660 ggtttctgtc tagactcattgtgtgataaa ggacataata atttttatca ccttcatcta 24720 atataggttt gtcaactctatattagttgt tttcttgaag gctggttttc ttccaaaatt 24780 cagtcttatt ttcagtctacactagctttt aaatatactg tcctttagat gctttatcta 24840 acctcaaatt tctaatggatttgtcttaga cacttattgc cactccttag atagtcattg 24900 ctatctttga agttctggacgatacgtgta ttacagagga actggagaca ttccatcacc 24960 atagttagct tgattggataccctttaaaa gcatatactc gcgcctgtaa tcccagcact 25020 ttgggaggcc gaggcgggtggatcacttga ggtctggagt ttgagacaag cctggccaac 25080 atggtgaaac ctgtctgtactaaaaataca aaaattagcc tggcatggta ccacatgcct 25140 gtaatcccag ctactcaggaggctgaggca ggagaattgc ttgaacctgg gaagtggagg 25200 ttgcagtgaa ccaagatcttgccattgcac tccagcctgg gtgacaagag cagaactcca 25260 ttaaaaaaaa aaaaaagcatatatagcaca tattataagg ttttcaattt tttcaccaag 25320 tgtttcattt gggtagtcatttattggtag tttacatcag ttgagtggtt cagaaaaaat 25380 acagtaagtt gcttataaaattctgaacac tttggccagg cacaatggct caagcctgta 25440 atttgagccc tttgtgaggctgaggcagga gaattgcttg agcttaggcg ttcaagacca 25500 gcctaggtaa caaagaacgcctggaatgat tgtggcattt gaactaatat tcaggtttaa 25560 caagagataa ttgaccatcactctatttta gaggctttat ttgaaccaga tagaaatcta 25620 tttcccacag ctatcactgcctgtcaccta caacttaagg gggttgggga ggaagtgaga 25680 gattttctgt tagggccaatagggacctgc tagatacccc cccatcctgg gaatggtgta 25740 tggaactcca gtgtatgctggagttattat catcatactt gtttttttat tttactcttc 25800 tgcttataca gatcaagtcttacgttttat ttttaagttt aaattgaaaa catttacaga 25860 gaacaatgca gtgaaatgaaaaaattacag actgctggca tttgcatttt catgtagcct 25920 cagtgactaa tttttttttattgtacagca ttgagaaaat cctagttcat ataactagtt 25980 atagttcata tagattcatataactagttt taagtgataa tagtttcttc ctttttttcc 26040 tccaccatct aaccagatgaagataatagt ttttaatagc tcaccgtaaa tttcaaggta 26100 ctcaagttaa attgatctagatgcttgagt tgaaattttt ctatcaaagt tcaataacat 26160 gcttacattc cttattaaagtataaaagtc ctataaacac acaaacttga gtaagtacta 26220 aaactagtat cagtattgtcacaatacaac atgttatatt gtaacaagag catttgctga 26280 gaactgtgct tgttactccagaatgttgct tctatggttg tacctttcaa ctttgcagat 26340 catttggaag gaggagagatttggggtgga gacaattcgg tacttcattc acaggatgta 26400 aggaggatta agtaaaataatgctggctaa aagtccttat ttagcatact gcccaatgct 26460 cactaaatca taatagctgtttttaacatt tggtgaagaa tctatttaac aggagtgagt 26520 tgaggggcat aggagatcatgtgagtgttt aaagtagaag cagcattccc cattaagaag 26580 agaaatactg tggaagagcaaagactttaa aacacctggg ttcaaatcct atttgctaca 26640 taatggctac ttttaacctattgaacgcca gttccctcat ttgtaaaata gggacaatat 26700 ttaacctatt tacaggttgtgagagaacta ggcacctagt acagggtaat gttggcacat 26760 ggtaaccttt aataaactgttgctattcaa caagctatta gatgtcacta ggcagttaag 26820 caaaggaaga cagcttttgcttggtgtgac aatgaaaatc tttctgattt ccttcttgga 26880 agagttccct gaagatatgtcattgtattg acacctttat ttttgctaac ctatccctct 26940 aaattctgga tattgtgtgtgccacagctt tttttcttcc atattcctgc atttatttgg 27000 cacctgttgt gccagtaatagataaggggc tgctaaggga ggaggcaacc tgcactggct 27060 tatagctgct aatgtcagttcctatagctt atcgtcagtg ttattcatgt ggtaaaaggg 27120 tgagaaagta ctggagtctaaagaaacaag tagaaatcag tttgtagcta ttaccgttct 27180 acctgctaac aactcctgttttcaagttat tatgtacaac tttaggtagt ttctctagcc 27240 ttaatcgtgg tttctctgtattgagactac ttttgaattc tatgaagtac agccttagat 27300 gtacaggcta ctttaaatttttgcctaaaa taaaaacatt ctctccaatt acatatgctg 27360 gggaggaaac acctgcttccgacaggttta aagcttggtt ttggactttt tgtgagagtt 27420 ccttatgtgt gcagtaatccaaaatttgta tagttgccct ttataaaagt acattaatct 27480 agtagacaaa tctccatgtaacttaattac atggcatctt ctaatccttc tgtgataagc 27540 agaaatgtaa agttttattcaagttaaggc aaactaactt gtatacactt tccatctcgt 27600 gtttttcttg ttgttgttaagtaggataag ttctgaacgt cgaaaagaaa agtctcgaga 27660 tgcagccaga tctcggcgaagtaaagaatc tgaagttttt tatgagcttg ctcatcagtt 27720 gccacttcca cataatgtgagttcgcatct tgataaggcc tctgtgatga ggcttaccat 27780 cagctatttg cgtgtgaggaaacttctgga tgctggtgag ttattttaca agggtataaa 27840 taggcctgaa aattagaagttagaagtaaa tagaaattat ttttagaagg tggtcgcaat 27900 gttttgattt tgtatacctctttatattgt gatatgtaca cgtttaaaaa tttttctgta 27960 attctcacta tttttatcaagcttcatttt tttctcatca gttattcttt gaaataatca 28020 ttctttatgc acataatttgttttgcttta ttctcttaaa catactctca attcttttct 28080 aatataacat cctttttattacctgctttt aaagctttag tcaggaataa gatactggct 28140 tttcccctcc cccctttttctcctgttcca tctacctttc ttcctttaaa aaacatgact 28200 caggccgggc gcggtggctcacgcctgtaa tcccagaact ttgggatgct gaggcgggtg 28260 gatcatgagg tcaggagttcaagaccagcc tggccaagat ggtgaaaccc catatatacc 28320 aaaaatataa aaaattagatgggcacgctg gtaggtgcct gtaatctcag ctactaggga 28380 ggctgaggca ggagaattgcttaaactcag agggcggagc ttgcagtaag ccgagatcaa 28440 gccactgcac tccagcctgggcggcagagt gagactccat ctcaaaaata ataaaataaa 28500 taaataaata aaaaacattactcttctttc ttcttctatg gtttgctttg ctgcattact 28560 ttaatcatga aaagcagctggcacatctaa ttatagtttt tctagcttct ggcctgcact 28620 tttctgtgtt gaaatggctgtatatattaa ataaagtgtc tgcgagaaaa ctttgtaaaa 28680 acatctaaat attatatcatttaagtacaa ctttttaact aattattttc ctcttcttgt 28740 gcccttttta ggtgatttggatattgaaga tgacatgaaa gcacagatga attgctttta 28800 tttgaaagcc ttggatggttttgttatggt tctcacagat gatggtgaca tgatttacat 28860 ttctgataat gtgaacaaatacatgggatt aactcaggta aaatgcacac atattaagag 28920 ctcttctata tgtttttatgattttatgat ctagccctaa tttttaaaaa tgtgtttaca 28980 gtttgaacta actggacacagtgtgtttga ttttactcat ccatgtgacc atgaggaaat 29040 gagagaaatg cttacacacagaaatggtaa gaaaagtctg ttgtttgatt taatgtgaca 29100 ggtggtttta cataataagatactattgct aattattaaa ctttgctatt gtacttaccc 29160 aaggcaaaat gttatttcatgtttaataaa atgtctattc tttgttaaaa ctattatttt 29220 agtttttagg aatttcattttgaaagccca cctaattgca taaataattg tgtgggtgtg 29280 agaaataaaa tggaaaagtaaaatcatgac caagagagtt acaaataact tttttttttt 29340 ttttttaaga tggggtctcgctcttttgcc catgctggag tgcagtggca caatcagctg 29400 actgcagcct tgaccgctgggactcaagcg atcctcccac ctcagtctcc caagttagct 29460 gggaccacag acgcgtgctaccatgcccag ctaaattttt aaaaattatt tgtagagaca 29520 aagtctcact atgctgctcaggctggtctt gaactactgg gcttaagcca tcctctcacc 29580 tcggcctctc aaagtgttgggattacaggc atgagccacc acgcccaggc tacctttttt 29640 ttccttttct ttttaaattgtgataggggt tcttgctgta ttgcccaggc tggtcttaaa 29700 ctcctggact caagtgatcctcctggctca gcctcccaaa gtgctaggat tataggcatg 29760 cgccaccaca cctggtggagttaaaaatta aaatacacca ttaaggcaag gagaaattat 29820 aatacaaatg gcagataataggactttaga cagtcattaa agttgaggtg ccagtttgag 29880 tctaaggccc aataaaaaaagttcaccaga attttaagac aaacaactgc ttatttgact 29940 tctttggatg ttctcaataattcgagaccg tgtagttaga ttataaagta ttacattgtg 30000 gatgcccaca tattaacaaaaatagagagt aagacctcta attcttagga attaattgtt 30060 aaaaataatc aagtgttccaagattttttg gaaactacct cttgaattaa aaaattaaag 30120 tctttctaca tttttatcttgttaaacagt gtatactgat cataattatt taaaaaatca 30180 tgtgttctaa gatttttggaaagtacctct tgaattacaa aaacaagaaa gtctttccac 30240 atttgtgcct tcttaagcagtgtatactga tcataattga acttttcttc atgatggaaa 30300 gttaccacaa ggaaaatttcttatgttctg ctgttctttg ttgctctcca atttaagtgc 30360 atacgtttgt ttgcttctatattataaaac ctcaaattta ctttttgtat aatttttgag 30420 gttttctttt tcatctcatttattataata atagctaacc tccattgaga gaatgctgtg 30480 tgccaggaca ctgttcttcctattttatat gcttttaact cctttattcc tcacaacaac 30540 cctgtgaagt taactgttagacaatttcta ttttactagg aaactgaggt acagagttac 30600 taagtaactt tcccaacattatttggttag taaatggcag agcttgggct gaacttcagt 30660 agactggctt cagagtccacgctcattagt cctttggagc gcttttcata ttcttgaatt 30720 ctcacattct gtcttttttcactctgtcag caggacctga ctcctgtttt taaatttcat 30780 attgtgtttt tactgttaatttggaaaaca aatgcatact ttttagaatt ctgtataaag 30840 gaggagtaaa tatgctgtgaacaaggacct aagtgggttg tcaatgagtt taatatatga 30900 gttctaatgt gcagagttgaggtttatatt gactgctcag tgcttccctg gggctagact 30960 ataaatggat ggatattaggaagtcttgtt ctgatttggt aatgatgtta atgcattatt 31020 ctaaatcaga tagtcttaatatagtttaaa tgtatgtttc gaaccaaatg ttctttttta 31080 aagcacacaa acattttgaaatcattacta atgtggttaa tgaattattg atgttccatt 31140 gggaaactaa aatgcagatttttctctttt agaaatcagg gactattgca aagcatcaca 31200 ttttagtgat acactgagagccagtggtgt gtttatacaa atagtcctat tttccaaata 31260 aattctagaa aaatgctttagaatttataa attatacaaa atatgactta tttttagaga 31320 gtttaaaatt taggtttttttaatggtttg tttttgtttg tttgtttttt gttttttttt 31380 tcctcattag gaaaacactagtacttttca gttaccttga tttttaaatt aatctgcagg 31440 tccccattca aaggccttgggttcctttca aaggtcagta taattcaagc ttagtttatg 31500 aaggactgaa catacccaaaggattttgca tgtggatctt tactgccact accacaacca 31560 tcaacaccta cacacacacgacacacacac attctctctc tctctctctc tctctctctc 31620 tctccccctc cctcccgcactccttccctt ccccctcctt tgctctcatg gcatctttta 31680 aaaatatact cttaaatccttccagggagg gcaaattcac ttcttaatct aagtaaaccc 31740 aaatggcatg catcagcaccaggactgccc atctttccta gttccattat tcatagagta 31800 taggctggaa ttcatcttgttcctcaagag tccagcattt ctagttaacc atgcctacat 31860 ttaaacttac tctcatttcttttctacttt acagtgtttt ttcaatatac tagcattaca 31920 gtttccagat ttgatttctctcctgtctta tttccatcag ttttcaagtc tattaagatt 31980 ctacctcttc atttgtcttttgccaccatt cttttccctc atactctact ggctcagccc 32040 tctcattaca gtcacctaattctaacatat atattgctgc taagttaatt ttccttaagt 32100 tactgattgt gcttttttaaagccccttgt tgaatattta ggcaggactc catgtggaca 32160 tccacagccc tccgtggtacagccctaacc ttcccttcta gctttgcctt actactcttc 32220 tacgtgtact ctacattgtggacaaactac tatatgctgt ttttcaaaca tgtcctattt 32280 ttcctacctc tgtgcttttcattctcttac ttctccttgg aatacccttc taacccatct 32340 ctacttactg acattctaatgtctcttttt ctaagcaaga cttcttgatt tcccttgact 32400 agaaattatc ttctaagctctccctatcct tctttaaagc atttttataa gtctcaagta 32460 ccaactctac attgtgtttttgttgacctt actatatcta ctacattttt aacttcttca 32520 ggaaaggtgg cgtatcttactcatctttgt attgcctaca atatctagtc caggttctga 32580 ataataaata tttttatatgtgttctgaag cacactgacc aatgaagata agaaatcaag 32640 aggctagttc cttattttttttaatttttt tttttgagac agtgtctcac tttgtcaccc 32700 aggctggagt gcagtggcacaatctcagtt cactacaacc tctgcctccc gggttcaagt 32760 gattctcacg cctcaacctcccaagtagct gggattatag gcatgtgcca ccacacctag 32820 ctgatattta tatttttagtagagatgggg ttttgccatg atggccagca tggtctcaaa 32880 cttctgtcct caagtgatcttcctgcctca gcctcccaaa gtgctgggat tacaggcatg 32940 aggcataagc cactgcgcccagcaagatgc tcttttctca gtcacctaaa tataatctca 33000 tttttagtta tagaaggtttgaaattggag tgaatagact ttacttaatt ctgactttat 33060 ttctgtagct ttttttttttgagatggatt ctcgctctat atcccaggtt ggagtgcagt 33120 ggcacagtct cagctcactgcaacctctgc ctcccacgtt cgagtgattc ccctgcctca 33180 gtctcccaag tagctgggattacaggcacc cactatcaca cccagctaat ttttgtattt 33240 ttagtagaga cagggtttcaccatgttggc caggccggtt tcgaactcct gacctcaagt 33300 gatcctcttg cctcagcctcccaaagtgct gggattacag gcatgagcca ccgtgccctg 33360 cctatttctg taacttttgataagtcattt gatctgttgt tgttgttttc tcatagtaac 33420 aaagtagaag taattttctgcctgctttac tagataaatt aaggggaaaa aaataagata 33480 cgtaaaaatg ttatttgttattaaaaagaa agttgttatt ttaaaggttc tataaagaca 33540 tagagtgctt attagaaattgagctaacac attcaggaaa ggataggaag agtttgctga 33600 agttctttct ttagggattcttgtgtaccg atagcacagt taaagagcaa actcatacca 33660 tttttatatt tctgtgtatttgactaagct tactggcttc aatgattaac tgttatccca 33720 aatatggatt atctttcagccaactcaggg aatcacagct actgagtagt gtgtgtcaga 33780 tctcttgggt gtgctggagtgagtaaaagg ggaatgaatt actgtgttca tgctgagact 33840 taattgaacg ggtattcagttgatctaggt gatgggcact ttgttacttt tattgtaaca 33900 aatttgtata tttagttgctttaaaacttt atttcatgct ttcattaggc cttgtgaaaa 33960 agggtaaaga acaaaacacacagcgaagct tttttctcag aatgaagtgt accctaacta 34020 gccgaggaag aactatgaacataaagtctg caacatggaa ggtaagtgaa aattatttgt 34080 gattgattat acactttatttatacataga cattgtagta ttaagataac tttagaattg 34140 tgagggaagg tttacagttccatggtgttt ggttatgtaa catttatatc ttcaactcat 34200 ttgcatgtga tctccaaaatgcagaaccgt gtagtaattt gccaatttga ggcacaaact 34260 taaattacgt gaattgtggcactggtgttc caggcttaat cagttggctt tgccagccac 34320 acaatatttg aatcctgatagggcttaatt ttctattaat catggtttta tatctttgtt 34380 caatgttgaa acatagtcatcagtgcaaga aataactatc aaacagccat gatgatgaga 34440 tgaatgaaaa agcagcctagactttatacg aggggaattt tttaaagagt aatgtatagg 34500 ccctgggcag gaagtaggtcataggtggta tcataggaaa aatgttcatt gattttcaaa 34560 aacgtgatta atccactagtgacagtaaat tttatcaaag cttactggcc atgtcagact 34620 caactactta tctctgctttttttttccct agcattgtaa atattttttt taactgcttt 34680 gttcttcata cacaggtattgcactgcaca ggccacattc acgtatatga taccaacagt 34740 aaccaacctc agtgtgggtataagaaacca cctatgacct gcttggtgct gatttgtgaa 34800 cccattcctc acccatcaaatattgaaatt cctttagata gcaagacttt cctcagtcga 34860 cacagcctgg atatgaaattttcttattgt gatgaaaggt aaattagatc taaaatgtga 34920 atttgaaatt tttaattagtctacagcatt actgaatatt caccatagca aagattcagc 34980 gctggccatg catggtggctcacacctgta atcccagcac tttggaaggc tgaggcaagc 35040 ggggggtgga tcatctgaggtcaggagatt gagaccagcc tggccaatgt ggtgaaaccc 35100 catctctact aaaaaatacaaaaattagtg ggacgtggtg gcaggcacta ctcaggaggc 35160 tgaggcagga gaatcgcttgaacctgggag gtggatgttg tggtgagctg agctcacacc 35220 accacactgc aagcctggatgacagagcaa gactcccatt tcaaaaaaaa aaaaaaaaat 35280 tactcaatgt taaactatactttccactaa attgaacaga atgatacatc ctataatatt 35340 agattaactt tgtaaattaattcagccaca tttattgaac atttactctg tactatgaac 35400 acttacttta ctaggtgctatccagaagtt aagatgagtc tttttttccc caataggggc 35460 tctacttact tagagaatttcaaagatatg cagtgtgtat tttgagcaaa gatagattac 35520 cttaggttgg ggactagaaagccaagtgtt tgtacatctc ttcatcctac atattttccc 35580 tgagaagctt caaccttgcccatggtttct attactattt cccacatttc ttcctgtaac 35640 taattctatt taattgccaacttaatattt ctatctggat attcttctgt attgtaaact 35700 aagtattact gtaacaactgtactactact gcccccaaac aacatcatca tcaaaaactg 35760 cctttcttcc tataatgcttattgtggttt aatacaccac catacacaca tgactccagc 35820 aaaactttgg aagtcatctgtaacttttct tttacattca ttggctacat acagttggtg 35880 tctaaatctt acagatttactatctacata tatctcttga tccatttcct cctttccatc 35940 cttgcactcc tgccattgaattcattagct cattattact cttgacttga gttgttggca 36000 tagctgcctt tttgccaacagatttgtacc cttataatct ttcatctaag ttgccagaaa 36060 gtgggtgtcc taatgtgaaaatcagatcat gtcattctgt tgttgaaaat gcctcaaatg 36120 cttccctcca tctttgcacacaaaaatatt ttgtttataa aaatactaga tgagggaagt 36180 aaatttttca tttatcaaaagaagatgtgt attttagaag actgaaaaaa aatagaccta 36240 cacaatacaa tctaaacttagcatggcaaa caaagatatt tatgctctgg ccctaactct 36300 gtctttggaa tcagatgttagattcactca tggcttgcag ctctgatact tacaatgtgg 36360 ccttggcctt ggtacttaactgttgtaaaa ttcacattcc ttatctataa aataagaatc 36420 atggctgggt ggggtggctcatgcctataa tcctagcact gtgggaggcc gaggtgggtg 36480 gatcacctga ggtcaggagtttgaaaccag cctggccaac atggtaaaac cccatctcta 36540 ctaaaaatac aaaaattagctgggtatggg ggcacatgtc cgtaatccca gctacttggg 36600 aggctgaggt aggagaattgcttgaatcca ggaggcggag gttgcagtga accaagcttg 36660 caccactgca ctccggcctgggagacggag tgagactcca tctcaaaaaa caaaaacaaa 36720 acaaaaaaaa gacctcagaaggatgttgtc aggattaaag gagtccattg agtgcctagt 36780 acagatagtg aatgcttcactactggtgtc aactttaaga aaatgaatat agaaaagcta 36840 agaattattt taaggtgtttactactagca tgtaaatgta tgatgggaca gagatttcca 36900 tcctattttg aggaattattttttattttt ttgaaaactt aaggtaacaa agtagagagg 36960 aggccaggga gaaaggaaggtagtggagca aaaatgagaa agggagtgac attcccctct 37020 agttatagca gaaaattagcaaaatgatca tgacaggagg taacagtaaa gacagccagc 37080 tcatatatca accaagacagttttgagttt gaccagcaga ctgttatttt ctggtttaga 37140 gctctttcca ggaacttcttgcatctataa cccctgagaa ccaagctatg gaaaaaattt 37200 tgctcaattt taagaaaatctaacatatca agctcctcaa ctccaaaata ttccacaaat 37260 agctgctatt tactatactgagtaataatc atttaaaatt attcaacact ttatttgagc 37320 atctactatg ttcatggcactaaagtagaa atgaagatga acagttcctg cctcaaaata 37380 aatgagtagt atactgctttagatcatggg tttcctagtc cattaaaaac actttttggt 37440 catattttct ggacaccccgacccttttgg tatagaatat aacctatgta attctctaaa 37500 gttaaattaa cctcacttttcttgctctaa tatgtgtaaa actgaccttc taggaaagca 37560 tatacagttt atatttttgacttcttggta tcttttagtg atagacatac ctcagattga 37620 gaagcactga ttgacattagattaaatcag agcttcctat gacaatataa acaatacctt 37680 cattaatctg atccccctacctacttcttc agcatcatct catatctgtc tccactaatc 37740 atattataga atctttgttacctgcaccat gttaagcatt tttaaaaatc ttttgtttat 37800 accatacctt tttcctgaaagcggttttgc ctttcctttg tctctagtca taagtctcct 37860 ataagaggct gttcctcattctaccattcc tttgcatgga taggattcca tggaatagat 37920 tctcatcact gcatttatcacattatttcc taagtagtac agtacatcta ctggaagatt 37980 agccacgtat tgagttttgtctttgcattt tcatgcctag aataatgccg ggcacacata 38040 ggcatattaa gatttgaatagtgaaaaagt ttttaattcc atggggattt tatttaaaca 38100 gaaaaatata agaccaattagaattatttt taaagcataa tttcaagaaa tatgactgat 38160 tttgtttaaa aacatgttttcctttataat gctgccacct ggtgttgctg tgtttagaga 38220 tgtccctttg taaagaattgagggtttgag ttgagtttgg tttggttttt ggcaaatcag 38280 cttttccttt gtatatttattttgtaataa actatggaag atcttgcctt taagtgtgag 38340 aacacaagca atgttacttttataccttta tagaatatct tgcctatgtc cttcctgtag 38400 ttaggtaggg ttttttttttgacacacagc atgttatata aggtttgctt gcacctcggt 38460 aggaaagtcc tctgaaatctaaaggctgag aatctaaaag cttaactcat gttttgctcc 38520 tagaaagact tgagaagagagtatttctgt tcagcatggt actaagaaga cagctttctc 38580 ttcctcatgt catggttgccatttcatact gcttacagag aataagatct agtctctgtc 38640 ttaaataaag gtctactctctgccagcgag ctagataggg taattggatt gttttccaat 38700 ctattttcat ttgaaatattgttttatctg aaattactcc cataatttca tgtaatgcca 38760 aaaactaaac taagtacaagagcatcttca aaaaccaaca taattccttt agttcccatt 38820 tagtgtagat gctctttggttgatgatatt agaattgtgt aatggctatt gatctctcaa 38880 agtgaggtgt tgcctaggggcttaaaagtt actacataaa gaatttggct ttatgaagaa 38940 atgttacaga ttttatctatattttaaaat aagtgtaagt gactaccttt ataactttta 39000 ccatgtagtt tagtagtatttcttatctgt ttattaatac cctgccttgt taccaaaagt 39060 atgtataatg agatgtaataagaataggta acaagtaggc tgggcacgtt ggctcatgcc 39120 tgtaatccca gtactttgggaggccaaggc gggtgaatta cctgagttca ggagttcaag 39180 acgagcctga ccaacatggagaaaccccat ccctactaaa aatacaaaat tagctgggca 39240 tggtggcaca tgcctgtaatcccagctact tgggaggctg aggcagggga atcgcttgaa 39300 cctaggaggt ggaggttgcggtgagccaag atcacacctc attgtgctct ccagcctgag 39360 caacacgagg gaaactcttgtctcaaaaaa aaagaccagg taacaagttt gggtgaacag 39420 gattaaagag ttaaataacaggaggaatct agaggactta aagaaatgtg tggtgttgga 39480 tttaataact gtagttgccaaaggtgaggt gtaaatttat tctaagcaaa ggaggatgct 39540 catttttgaa aattcacttgtccataagat taatgcctat cagttaactt gggaggagaa 39600 aaatttttct ttatcagtgtctcccttttt tttcttaaat cttgtatttt ttactaacag 39660 aattaccgaa ttgatgggatatgagccaga agaactttta ggccgctcaa tttatgaata 39720 ttatcatgct ttggactctgatcatctgac caaaactcat catgatagta agtacaatgg 39780 aagaactcag agatattctaattacttaac tgttgcaacc tctgtacagt ttggctaccc 39840 atctaattct ctggttaaaagttctagact aaatgtgtta acaggcctat tcagtagaga 39900 tcttgaccat tttgtgttttgtatgtgttg caacaaatat cagtaaaaat agaatcattt 39960 aatcatagaa aaaacttcctggcattttaa atacaaagac ttttgaaaat ccaaatatta 40020 tagagtattg aatagcataattttcagaat tcacataaat actcagaaca gtggttggta 40080 tgtaaaaggc actcagaaagtatttgtaca atcaatgaat gtgaaggtgg tgaacatcac 40140 ctttggtaat aagtaccattttaaaaaatg cttataagtg catagttagg tatttatatt 40200 tatgggttca tgaaatattttgatataggc atgcagtgca taaggataaa tggagtacct 40260 atcacctcaa gcattatcttgtgtgacaaa caatccagtt atactctttt ggttattttt 40320 attttatttt attttatttttttcttttga gacaggatct cactctcgcc caggctggag 40380 tgcagtggag caatctcagctcactgcaac ccccgcctac cgggttcaag agattctcct 40440 gcctcatcct cccaagtagctgggattata agcatgtacc accatgcctg gctaattttt 40500 gtatttttag tatagacagggttttgccat gttggccagg ctggtctcga actcctgacc 40560 tcaggttatc cacctgccttggcacccggc ctcttttagt ttctttaaaa tgtacaatta 40620 aattattttt tactatagtcacccaaaaca agtacctttg acataagatt tgattctgaa 40680 ttttactcaa atgaatgttaagatccccaa gataagttaa actttggact atctcacctg 40740 tttaatctgt acctatgcatgacttcccac tgtgcttgag gatacctgaa tatcactgag 40800 tttgtgtgac tgatcagccttgaactcaag agtaaatcca agtctgcagt caggacaccc 40860 caatcctcaa aataataccatcattagcat ttatttagta ctttctccca aatcagtatt 40920 taatttaaat tgccaaaagacttacaatgt ggtatcaatt tatatttaaa tatgctacat 40980 atagcttttt aaagcatctttggttctctg gaaaccatag tcagaattta aggaagttat 41040 tgtggcacca ttttcttgaaaaaggctatt gattattctc taatctgaca ccaacctaag 41100 tcattaaagg aattttagttactgaagatt gtatattcat gaactcttca cttagctcac 41160 tggcagcaaa ggagttttatttagggggtt tgaaaaagga aatgggtaca ttttcagcta 41220 ttctgggacg cactgtcagaatgtaagcag ttacaactga ttccactaaa taaacatttg 41280 ttttccaaaa caatgatgaacattcagcat ctgttcattt aattgaaaat tcaaagttaa 41340 aatattttct ctgcatgattctttttcttt tcccccctag tgtttactaa aggacaagtc 41400 accacaggac agtacaggatgcttgccaaa agaggtggat atgtctgggt tgaaactcaa 41460 gcaactgtca tatataacaccaagaattct caaccacagt gcattgtatg tgtgaattac 41520 gttgtgaggt aagtaagtttgagaaataaa catttttggg gaacaaatag taattctttt 41580 tggatactct gttcatttataggaagataa gataataaat attaactaaa ttttaattct 41640 tttacatcgc taccaaattattattttcta tactctgacc taggtttcca gtccagctat 41700 tccacagtga tgctgctaaacactgtcagt agttgtctat ccccatacct tcactcctat 41760 ttttaaaaag accatgaaaaaaataccaga tccattgatt ggtttggtct aattatacag 41820 atatcggcat atactatctcaagacagctg tgttcttttt gtaggaagaa tcctggccta 41880 gatttgtatc atagctctaccactcattag ctccctgacc ttggggaagt ctcttcattt 41940 ttctgaattt catctatgtagataatcctt cagaaggtta taatgaaaat taaatgaaat 42000 tctatgagat tagggaggggggagggatag cattaggaga tatacttaat gtgaatgatg 42060 agttaatgga tgtagcacaccaacatggca catgtataca tatgtaacaa acctgcacgt 42120 tgtgcacatg caccctagaacttaaagtat aatttaaaaa agaaaagaaa ttctatgaga 42180 ttaataagct atatgatgtaatacatggct cttgtatatt catgaactct tcacttagct 42240 ctttggcttg tgaatattatgtacatcaaa atttaatttt tcatttgatc tattttacta 42300 gactcctgcc ccatctagtctacctgtcca cattattacc acattctagt ccatcttgcc 42360 cattactacc aggctaagctttctagtgtg gatatgtcat catcttattt tccttagaat 42420 tttagcgatc tttttatcatttccaagata aacacttgcc taggtgtaca gcatccttgt 42480 ttaccatcat actcacgcattagagattta gccttccctt taaaatctag ggtcactcct 42540 cttaggaaga ctttgggcagtttttatttt tgctacttct gacaccatcc tttaatgttt 42600 taatattagt gccacagagttcttttgtga ctttaccatt atgtaagaat cttccacttg 42660 gaatgtcttt ctcttcctcacaccccagtc tgcctagcaa atgccacttg atcccaagta 42720 tcagcttgtt agcttctcagtgaagcaagc cttctctatt ttagcagtta tcacagtgta 42780 ttttaattgt ttacatatctactttcacaa tgggttataa atttcttaag gtcaagggtt 42840 ggctatttta atctttgcattatcagttca tttcagatag tgaacattta atacgttaat 42900 taaaggaata atttacatttaagccaaacg tgaagataaa ctattgctca tcatccctct 42960 tcagccgtat cctgtaggtggtatcacctt atattcttac caccaaagaa aatatggccc 43020 ctctcttaga aagatcttaatcatttatct gtgtatcttt aggactatcc ttagatcatg 43080 cctcacatat tgatgccaaagagttctttt gtgccaattt cataatgtgt gtcagcacaa 43140 caattctgaa gatttgttggtgtctttcat gtacttgact acaaattgcc ttgccattac 43200 tactcttctc aaaggatatctgaaattctt tttttctttt ttttttttga gatggagtct 43260 cactgtcacc caggctggagtgcagtggcg tgatcttggc tcactccatt tcccgagctc 43320 aagtgattct catgcctcagcctcccaagt agctgggact acaggtgtgc accaccacac 43380 cgggctaatt ttttgtatttttagtagaga cagggttttg ccatgttggc caggctcttg 43440 aactcccagg ctcaagcgatccacccgcct cagcctccca aagtcctggg attacaggca 43500 tgagccacca cgcccagcctggatatctga aattcttaac tgaaattagt caaattatct 43560 tgtactgggg attttttttttaatttcaac ttttattttt gattcagggg atacatgcat 43620 aggtttgtta catgggtatatcatgtgatg ctgaggtttg gggtacaatt gatcctgtca 43680 cccaggtagt gagcataatacccaacagtt gttcaaccct tgcccctctc ccctagtagt 43740 cctcagtgtc tattgatgccatctttatgt ccacaagtaa cccagtgttt agctcccact 43800 tacaagtgag aacatgcagcatttggtttt ctgttcctgg gttatctcac ttaggataat 43860 ggtctctgga tgcatccatgttgctgcaaa ggacattatt tcattctttt ttatggttgc 43920 atactgtgga ttttattgggtctttatttt gtattagcat tttaaaaccc taaatgtgac 43980 acagtacgca tgagtgatcatgcatctcaa gaaatcttga aatgttcctg tccataaagc 44040 agaatttttt aagagaccatttcacagtct cccttcccct cactgtatca agtgctcatt 44100 tgtgaattac caatttctcttgttttgaca gtggtattat tcagcacgac ttgattttct 44160 cccttcaaca aacagaatgtgtccttaaac cggttgaatc ttcagatatg aaaatgactc 44220 agctattcac caaagttgaatcagaagata caagtagcct ctttgacaaa cttaagaagg 44280 aacctgatgc tttaactttgctggccccag ccgctggaga cacaatcata tctttagatt 44340 ttggcagcaa cggtgagtagttatttttgt taatccccta aattgtgtct gttgctacaa 44400 gccccatttc aactaaacattactttacgg tttttgttgg taatcatttg gacattacaa 44460 gctaatatat gtttatagttttcttaaatg tatttgctta aatatttttg cccccgtaat 44520 ttcttaccat tcttgcttttttatactgtt ggaaattgtg cttcaaagtg tccttaaggt 44580 atttcttctt cccacataaatttttcctgg ctactctatt tctgtatcct gctgtcagat 44640 tttctccaca gtttagcagagttatatgga agtaggcatt gttgcattaa aggataaaaa 44700 agtagtcata ctataacatcaagcattgaa gatgaaaact gcaattttaa agtagagaac 44760 attttaatgt ataaaaaggttggtattgcc ttttgtcttt tatgccatag agattaagac 44820 gcggtatcaa tagtggattgtaaaggtaac tcagacttat ggttatacta tactattgta 44880 tgtaaacttt ctgatgaaggaaaatttggt gacattttgt tgtttgatga attagacaaa 44940 ccttttgtga aaaagaacataaatttttta tatgtgaaaa tccttgtggc cgggcgcagt 45000 ggctcacgcc tgtaatcccagcactttggg aggccgaggc gggtggatca cttgaggtta 45060 ggagttcgag accagcctggccaccatggt gaaaccccgt ctctaccaaa aatacaaaag 45120 ttagctgggc gtggtggtgtgcgcctgtaa tcccagctac ttgggaggct gaggcagggg 45180 aattgcttga acctgggaggcagaggttgc agtgagccaa gattgcgcca ttgcactcca 45240 gcctgggcaa cagagcaagactctgtcttg ggtaaaaaaa aaaaaaaatc cttctatact 45300 ttagattgac tcatattttttccccacaga cacagaaact gatgaccagc aacttgagga 45360 agtaccatta tataatgatgtaatgctccc ctcacccaac gaaaaattac agaatataaa 45420 tttggcaatg tctccattacccaccgctga aacgccaaag ccacttcgaa gtagtgctga 45480 ccctgcactc aatcaagaagttgcattaaa attagaacca aatccagagt cactggaact 45540 ttcttttacc atgccccagattcaggatca gacacctagt ccttccgatg gaagcactag 45600 acaaagttca cctgaggtaggtgtcatgat ataatcagaa agggacaact ttcagatttt 45660 aacattcaag aatgtatttataagtttgat tcaaacactt atttgaacca caaattacat 45720 ttgtgtgtgt gtttgaattttagcacttta aaattattgc aagagctact gcctaaccta 45780 gacctgagca catgttttaggctcaaagat agtcaggaac atgggaagaa actagcttaa 45840 tataaaccaa aaggtgaaacgtacattgtt tctctattat ttatatcagt aggacaaaaa 45900 catcttgaat ttggacatttaaagagaata gtactaagtg tgctcaaggt agctacagcc 45960 tatacctgtt accccttttagtttgtttta ttgtgttttg ttttgttttg agaaagagtc 46020 tcactatcac ccaggctggagtgcagtggt gcaatcacag cctcaacctc ccaggctcaa 46080 atgattctcc cacctcagcctcccaagtag ctgggactac aggcctgcat caccatgcct 46140 ggctaatttt ttaacctttttttgtgtgtg tgtgtggagt tggggttctc actatgttgc 46200 tcaggctggt tttaaactcctgggctcaag cgatcctcct gccttggcct cccaaagtac 46260 taggattaca ggcgtgagctaccatgcctg gcccattacc cctttgagtt ggagaactgt 46320 ctggtagcaa tagacttacgagggtttaaa tgggaaagga ccttataaat tctttgccca 46380 atttagtcta atttccatcactattttgaa attttgggta agtataatat gaaaataaca 46440 agtgttacat aaaataaatacttagtaact ggtctttttt attctggatc tgtcttgata 46500 ttaattgtcc tatgaacacaaaaataatct ttaaaggcta ggctggccaa gacttagaga 46560 tatcacacag ggctctatttctaaatctag aatgattcca ttttagggct tcctacatct 46620 aaaaatatgc tcaggagtagggcaacttag atctgaacat tataacttga taaatgaggc 46680 ataaataagc tttaataagtggtaaataat tctacattag gtatttgttg aataaaactg 46740 acaagctaag agtaggggatttgacatctc acagccttgt gttgaatgaa tatatatcct 46800 atgctctggt tgcttaatttacccagaaaa aaaaatgttt gattcatctt ggtttttatc 46860 taacaaaagt aaatctaacaaaaacgttag aatgaggaaa gcaaaatttc ttgtttagaa 46920 tacacagcta tagttttttgttaaacttct tgcccagaac tcttaaaata gtaataatgt 46980 acattcgttc aggtatatgcaggtaaaata acttaggttt ctactcccac ccccgacagt 47040 aacagtgaga tttttaggtagctcagtcac cacaggagtg tgccttctca gttcaaaggt 47100 aaattccagt gaatgtagcatctagttaat tggtcaatta ggtaccattg tgggatgtga 47160 attaccaaat aggttttattctttagaata aggtgtttct tttcatctca attttgtaaa 47220 tgatgttata ttacatagtcagaaatatat atattggcaa aattagttac cagtataagc 47280 ttcaaaatgt cactattttcacaaattttt tttttttttt ttttttttga catggagtct 47340 cactctgtcg ccaggctggagtgcagtggc atgatcttgg ctcactgcaa cctctgcctc 47400 ccaggttcaa gtgattctcctgcctcagcc tcctgagtag ctgggattac aggcgtttgc 47460 caccatgcct agctaatttttgtattttta gtagagacga ggtttcacca tgttggccag 47520 gatggtctcg atctcttgacctcattatcc ctccaccttg gcttcccaaa gtgctgggat 47580 tacaggcgtg agccactgagcccggcctag ttaaataaaa tttgataaac acgatggact 47640 tggttgtgtg ttttctggtttttctgagat ctagtttgaa aattctgaca actagcaaag 47700 tatatggaag cttcttcaggaaatagtaaa catatttctt tttacagcct aatagtccca 47760 gtgaatattg tttttatgtggatagtgata tggtcaatga attcaagttg gaattggtag 47820 aaaaactttt tgctgaagacacagaagcaa agaacccatt ttctactcag gtatatgaac 47880 ttatttgttt tatattaaatttcattaatt tttagtctga agtgactttg agtttcactt 47940 gttttttatt tataaggtgtggccattgta aaaactcatg tatttgctgt tttaaaggac 48000 acagatttag acttggagatgttagctccc tatatcccaa tggatgatga cttccagtta 48060 cgttccttcg atcagttgtcaccattagaa agcagttccg caagccctga aagcgcaagt 48120 cctcaaagca cagttacagtattccagcag actcaaatac aagaacctac tgctaatgcc 48180 accactacca ctgccaccactgatgaatta aaaacagtga caaaagaccg tatggaagac 48240 attaaaatat tgattgcatctccatctcct acccacatac ataaagaaac tactagtgcc 48300 acatcatcac catatagagatactcaaagt cggacagcct caccaaacag agcaggaaaa 48360 ggagtcatag aacagacagaaaaatctcat ccaagaagcc ctaacgtgtt atctgtcgct 48420 ttgagtcaaa ggtatttatatgtaacattc aagttatagt tcttttatta tttttgagat 48480 aaatgtatgt gatagtacatgatttttaaa cttatagcaa actttctgat atatatgccc 48540 taacgcaaat tcttgagaactcaaaaaact ttctaaatta acctcatata ttttttcttt 48600 ttctttcttt tttttttttttgagacagag tctcgctttg tcgcccaggc tggagtgcaa 48660 tggcatggca ccatctcagctcacggcaac ctctgcctcc tgggtgcaag agattctcct 48720 gcctcagcct cccgagtagctgggattaca ggcatgcacc accacgcccg gctgattttt 48780 ttggtatttt tcatagagacagggtttctc cacgttggtc aggctggtct caaactcccg 48840 acttcaggtg atccgcctgcctcagcctcc gaaagagctg ggattacagg tgtgagccac 48900 catgcccgct cctattttttctaaaataat tataaattct aaaattacct atctaaatgg 48960 aggagggtct tctgacacctttaaaataaa atccagctca gtactgtaaa tgtgtttaca 49020 gaacttgttt aaagttcttacagttgttta aatcagacta gttaactacc ctcactactt 49080 agatgcttcc atttcttagagctctttttt aagcttatct gaagaaaagc ccttccaatt 49140 taagggttat ttccaattgcacattccaaa ttgagccttc catcttcagc attcaatata 49200 gatatttaca ggcccctcttttaaaatttt attatagtta acttgtatta aagttgcttt 49260 tatttttcat tacgtatttgtagaacatta gctatatata tattgcaggc tacataggtt 49320 ttcaaactgt acaacaggaatctaagcatg aattgttact tctatggagc tagttcaaac 49380 aaacatatgg acatgacccaatttttaagt tatactttct gtatataatt tgtaagggga 49440 tttcacatat tttaagtttgaggctatagc tagaagaaat taagttttat ctaataagtg 49500 tgtggaaaag ggaaatgattccttctctac tatgtctaga ctaagccaga tatcaatagc 49560 aataggaaag aaccactgtcgtagccagaa cacatagctt ttttccctgc ctaacattcc 49620 caccttgacc tagagtgctgggagaggtct tttccctaag cttggaaaag acattggggc 49680 tttagatgaa ctcagaagtactttacatta ctttatttac tgtgtcactt actcactttt 49740 gactctgagc tccacgagggcaatcacagt gtcttgggca ttttagtgat actaatactt 49800 agctcatgac ctaatgtgtagtacttcctc aataaatggt tgttgaggca gggcgcagtg 49860 gctcatcact gtaatctcagcactttggga ggctgaagcg ggtggatcac ctgaggccaa 49920 gagtttcaga ccagcctggccaacatggtg aaacccggtc tactaaaaat gcaaaaatta 49980 gctgggcgtg gtggcacgtgcctgtaatcc cagctacttt gggaggttga ggcaggagaa 50040 ttgcttgaac cctggaggtggaggctgcag tgagccgaga tcgtgccatt gcactccagc 50100 ctgggcgaga agagtgaaactcggtttcaa aaaaaaaaaa aaaaaaaaaa gttgttggac 50160 tgacagatgc atgaatacagtagtaaaaat gacaatcact tataagttac agtttactat 50220 cagctacaga ggatgggatatccagttttc tgaacaactg ttctcttgta cttgtcaaag 50280 ccaaagtgta acaacacatcaagtcacttt agcaatttat ttttgagacg gagttttgct 50340 cttgttgccc aggctggagtgcaatggcgt gatctcggat cttggctcac cgcaactgcc 50400 gcctcgctgg ttcaagcaaatctcgtgcct cagcctcccg agtagctggg attacaggca 50460 tgcaccacca cacccagctaattttgtatt tttagtagag acagtgtttc tccaggttga 50520 tcaggctggt ctcaaactcccgacctcagg tgatccacct gcctcagcct cccaaagtgc 50580 tgggatgaca gttgtgagccactgtgccca gctagcaact gtttttaaac attagttcca 50640 atgtagtgta cactgaaaacttttatgaaa ggaatttcaa aaattaagat aaaccattaa 50700 aaacgtaatt actaagtactactactacta caatgatatt tacataatag actgagttac 50760 atttcataaa gacaatatatctgtataaga atttttaaac ttccctgtct atataataga 50820 agttttagag aaattttttaaaaaccaaag aaaactgcaa aataagatca cttacctatt 50880 tggcattctc aactgtctggaacagcaagg agccattatg attatgcatt tggtttgtgg 50940 ggtgtcttga aaagtcaaaataatgtaaca aagctgatgt actttactca ttagaacaat 51000 tcttcacaat ttaatattaattttagatat acatagttca tgtttgataa ccagatcaat 51060 actgagtgaa aaatagcatagtgggaagag caggggaggg gaggtaggga tctggagacc 51120 tagagtgtac ttccatattgcaactagtga gcagtaggac tttgagaaag ttacccaata 51180 ggcctcaggg ttctaatttataaaatgggt atgatatgcc tgccttatct gtcttgggaa 51240 cttaagtaag gttaaaatgaactaatgaac ttgaaatgtt ttataaactg aaaatgctat 51300 acgaatgtga gattgatcttgtatttcaat agtcccaaca atatcactgc attgttatat 51360 taggtggaat aaaaggacaatatttaactg ttttgactct acaatagtgt caatttagtt 51420 gtgttcagct ctattttataaaatagggat acgcatactg tagaaaattt cctgttaaat 51480 taagctttga cggccaggtgctcacgcctg taatcccagc actttgggag gccaaggtgg 51540 gcagatcact tgcgctcaggagtttgagac cagcctgagc aacatagtga aatcctgtct 51600 ctacaaaaat atgtatatataaattagtca taatcccagc tacttgagag gctgaggtgg 51660 gaggatcact tgattccagaggcagggctt ggttgcagta agcagagatc acgtctctgc 51720 actccagcct ggctgacagagtaagaccgt gtttcaccaa aaaaaaaaaa aaaaaattaa 51780 gcttttactt ttaagatgataaactttagt gatcaggaaa gttatcttat gtatattata 51840 ttccttaata ttggagaactaaagaattat gtattttctt taaaagcgct cactggatat 51900 tttttttaaa aacgctatattttcatttag aatttttttc ttttcagaac tacagttcct 51960 gaggaagaac taaatccaaagatactagct ttgcagaatg ctcagagaaa gcgaaaaatg 52020 gaacatgatg gttcactttttcaagcagta ggaattgtaa gtatgagtag taggttttgc 52080 ttttctagct aatgtgctatttcgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtttc 52140 cacgtttctt ccaaatagtaaagttatatt ttcagaagtt atacattggg tttttttact 52200 ctgtatgcac tggtttttaaaaatacaaat gtttaataca tacattcttg gtataaaaat 52260 tccaaacaat tccagtgtattttgagttaa aaagtgaagt tctcccctta ctccaccctg 52320 aatatcacca ccaatctcattctcttccct ttaagttact ttgccttatt aaaagaactg 52380 ctattggcca ggcacagtgcctcacgcctg taatcccagc actttgggag gccaagatga 52440 ggatcacttg aggtcaggagttcgagacca gcctggccaa cttggtgaaa ccctgtctct 52500 actaaaaata caaaaattagccaggcgtgt tggtgcacaa ctataatccc agccactctg 52560 gaggctgagg caggagaatagcttgaaccc gggaggtgga ggttgcgatg agctgagatc 52620 aggccactgc actccagcctgggtaagaga gtgagagtcc atctcatatt taaaaaagaa 52680 ctgctatgtt ttggggtaagtcaatggtgg tataatacat tctgatattt tcaaactaaa 52740 ttaactggaa agtatttatagacagaatgg tcataatgga tgacaaataa cttaagaaag 52800 aattcaaaat aatttagggtagtatttaag aaactgccta taatgttatt aaatttacac 52860 caatttcaag gtttttggttgtttaaaaaa aaaattcaac aaactaaact tgaaataact 52920 ttactgttta tagggaacattattacagca gccagacgat catgcagcta ctacatcact 52980 ttcttggaaa cgtgtaaaaggatgcaaatc tagtgaacag aatggaatgg agcaaaagac 53040 aattatttta ataccctctggttagtttat tctttttgac cttgaacatc acaaagacaa 53100 aatacatgaa acatttttatttaggagctt taatctaagt gagaatgact ttggttcctt 53160 agcaagatta aaaagtaaagttgtggctgg gcgcggtggc tcacacctgt aatcccagca 53220 ctttgggagg ccgaggcagccagatcatct gaggtcagga gttggagacc agcctggcca 53280 ccatggtgaa accccgtctctactaaaaat acaaaaatta gctgggcgtg gtggcgggcg 53340 cctgtaatcc cagctacttgggaggctgag gcatgagaat tgcttgaacc cggaaggcag 53400 aggttgcagt gagccaagatggcaccactg cactccagcc tgggcgacaa gggtgagact 53460 ctgcctcaaa aaaaaaaaaaaaaaaaagta cagttgtatt tcatgtgatg gtcttaatac 53520 agagattaac atttcaaggtggagcttttc atttttagta attttctttg atttctctat 53580 gtccatgtgc tgtcaatattgatagaagct gaaatttgtg aacttttatg acttcttttt 53640 tttttttttt tttttttgagacagggtctc gctctgttgc ccaggcctgg agtgcagtgg 53700 catgatcata gctcactgcagtctcaaact cctgtgctca agctcaagca atcatcctac 53760 ctcagcctcc tgagtagctcgcactacaga catgcctcac cacacccggt tgctttttgt 53820 agagatgggg tctcactatgttgcctaggc tggtttcaaa ctcctggcct caagtgatcc 53880 tcctgcctca gcctgtgctaggattacagg catcagcttt gatgcccacc atatttatgc 53940 ctttttccaa attgttatttctttgtgcct ttattgtatc ctgtaaacat ttctgacaca 54000 gcaacagtat cactggattatacttacttt ttaacatagt tgtggttttg ccaggtaaac 54060 taaaaaccct tccagaattttgctttattt tctatgatac ctaacacatt gtgggtgttt 54120 aataaatatt cattgactagatgaatgtat acttaggtat ctcttttgtt tttcagattt 54180 agcatgtaga ctgctggggcaatcaatgga tgaaagtgga ttaccacagc tgaccagtta 54240 tgattgtgaa gttaatgctcctatacaagg cagcagaaac ctactgcagg gtgaagaatt 54300 actcagagct ttggatcaagttaactgagc tttttcttaa tttcattcct ttttttggac 54360 actggtggct cattacctaaagcagtctat ttatattttc tacatctaat tttagaagcc 54420 tggctacaat actgcacaaacttggttagt tcaattttga tcccctttct acttaattta 54480 cattaatgct cttttttagtatgttcttta atgctggatc acagacagct cattttctca 54540 gttttttggt atttaaaccattgcattgca gtagcatcat tttaaaaaat gcaccttttt 54600 atttatttat ttttggctagggagtttatc cctttttcga attattttta agaagatgcc 54660 aatataattt ttgtaagaaggcagtaacct ttcatcatga tcataggcag ttgaaaaatt 54720 tttacacctt ttttttcacattttacataa ataataatgc tttgccagca gtacgtggta 54780 gccacaattg cacaatatattttcttaaaa aataccagca gttactcatg gaatatattc 54840 tgcgtttata aaactagtttttaagaagaa attttttttg gcctatgaaa ttgttaaacc 54900 tggaacatga cattgttaatcatataataa tgattcttaa atgctgtatg gtttattatt 54960 taaatgggta aagccatttacataatatag aaagatatgc atatatctag aaggtatgtg 55020 gcatttattt ggataaaattctcaattcag agaaatcatc tgatgtttct atagtcactt 55080 tgccagctca aaagaaaacaataccctatg tagttgtgga agtttatgct aatattgtgt 55140 aactgatatt aaacctaaatgttctgccta ccctgttggt ataaagatat tttgagcaga 55200 ctgtaaacaa gaaaaaaaaaatcatgcatt cttagcaaaa ttgcctagta tgttaatttg 55260 ctcaaaatac aatgtttgattttatgcact ttgtcgctat taacatcctt tttttcatgt 55320 agatttcaat aattgagtaattttagaagc attattttag gaatatatag ttgtcacagt 55380 aaatatcttg ttttttctatgtacattgta caaatttttc attccttttg ctctttgtgg 55440 ttggatctaa cactaactgtattgttttgt tacatcaaat aaacatcttc tgtggaccag 55500 gcccctttga tcagcttttatgttcaaata ttaataatat ttgcttcaac acctccaact 55560 cataaaattg tttaccaacaatttaagcac ttatgaaaat tacatggtac tggttatttc 55620 tacatttatc ttagtgccatcaccttaatg tatgttgagt ccctaaatgt catgttaaat 55680 aataacaacc ataatatcccattgaaaaga gtatgttgtt agaaaagaaa catcattttt 55740 aagtttctga gcctattaaaatgctcaaac acaaaatatt agtattttta aaatatgaat 55800 gggatgagtg aagcagttctcagcattata gtcacaatgt tacaaaggct agagcttctc 55860 tgaagatttc taatctgttcccattaacag attaataaat ttagacttca aatgaataat 55920 ttgcccaagc tttaaaagtaatagatggca gaccaaaaat gtaagcttaa gtttcctgac 55980 tctaaagtca aacttagaacaaatttggtt tgtttttgtt ttaatgatac tgcgttttaa 56040 aacaaagtag ctttatcctttttctcctgt atttttcttt tacaaaatag ctgtatttct 56100 tttatactga taatctcatttttaaaaatc agacagtgta gaaagatatt ttttaaaaca 56160 gaaaaatcac tatgaatccctgcacctaca ggtacagaaa attattttta tgaacaaatt 56220 atgtaggaag tgccagagccttaggtcctt taccctgagg tatatatact gaacaaaagg 56280 aactgagcca cagatctcttaggtagctct ttttatctta caatggagga cagtgattac 56340 aattatatga aaattttggaacaaaagtta atactaagat tcagtgcaaa atttgggggg 56400 ggggggggca caggtatacttaagcacaaa cactgtgacc caaagtgctt caacatttag 56460 ttacagatag tagtatactagaagtggtat tttagaataa agtggttgct tagtattcac 56520 aggtcacaaa acaaaaaattattcttgtat agcaaattag cttcagttga aaactatttg 56580 taaaagcaga ttatgtaatgaccaggagtt caggaaaatg acttctgaaa gcattgagaa 56640 gggaaagcca cgttaaaggacagtacagct ggaaggaagc aagtacttac ccactgctca 56700 gtcactaaga caacaagctccttggagtgc tttaagctac ggaatagcag aactggccct 56760 tcccaatttt atgcaccgtcacaaatttct tcataatggt tttgtccaag gcttataacc 56820 caaccctggc aactataatccttactttat gaaacagctg tatttctttt atactcataa 56880 cccagaaaaa tgagaatgtatgttctgagt ataaaagaaa tgtagctatt ccataaaaat 56940 acaggagaaa aagaataaagctattttaat ttttttaatg cagtatcatt aaaaaacaaa 57000 ccaagtttgt tctaagtttgactttagagt caggagactt aagcttacat ttctggtctg 57060 ccatctatta cttttagagcttgggtaaat cttcatttga gatctaaatg ctatatatag 57120 ttcattcata gcagtaccagataagggagg agtatatcta tacagtatat agtcttgaag 57180 aagtgatcta aggctcggagcttttgaggt ggccatgagt gactccaaag tccatggagc 57240 taaccaccct gcagtgctagccaatccagt tgaacatacc cttttctcca ttgttaactg 57300 tttgtttaaa tagcaaacagaaggcggcaa tggaggtgtg gaaaactgag gatccgatgt 57360 cacttgaaag taatgagatcacataacatt gagggaatgt cctaagagga gtggcagggc 57420 ataaatagaa atgaataaaagtgttttcaa gtgccattta gtgggttctg aatttgaact 57480 agagattgag atatccagtt57500 <210> SEQ ID NO 12 <211> LENGTH: 754 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (92)..(92) <223> OTHER INFORMATION: n is a, c, g, or t <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (181)..(181) <223>OTHER INFORMATION: n is a, c, g, or t <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (559)..(559) <223> OTHER INFORMATION: n isa, c, g, or t <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (598)..(598) <223> OTHER INFORMATION: n is a, c, g, or t <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (628)..(628) <223>OTHER INFORMATION: n is a, c, g, or t <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (678)..(678) <223> OTHER INFORMATION: n isa, c, g, or t <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (687)..(687) <223> OTHER INFORMATION: n is a, c, g, or t <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (694)..(694) <223>OTHER INFORMATION: n is a, c, g, or t <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (718)..(718) <223> OTHER INFORMATION: n isa, c, g, or t <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (748)..(748) <223> OTHER INFORMATION: n is a, c, g, or t <300>PUBLICATION INFORMATION: <308> DATABASE ACCESSION NUMBER: AU123241 <309>DATABASE ENTRY DATE: 2000-10-23 <313> RELEVANT RESIDUES: (1)..(754)<400> SEQUENCE: 12 ccctgacgct gcctcagctc ctcagtgcac agtgctgcctcgtctgaggg gacaggagga 60 tcaccctctt cgtcgcttcg gccagtgtgt cnggctgggccctgacaagc cacctgagga 120 gaggctcgga gccgggcccg gaccccggcg attgccgcccgcttctctct agtctcacga 180 ngggtttccc gcctcgcacc cccacctctg gacttgcctttccttctctt ctccgcgtgt 240 ggagggagcc agcgcttatg ccggagcgag cctgggggccgcccgccgtg aagacatcgc 300 ggggaccgat tcaccatgga gggcgccggc ggcgcgaacgacaagaaaaa gataagttct 360 gaacgtcgaa aagaaaagtc tcgagatgca gccagatctcggcgaagtaa agaatctgaa 420 gttttttatg agcttgctca tcagttgcca cttccacataatgtgagttc gcatcttgat 480 aaggcctctg tgatgaggct taccatcagc tatttgcgtgtgaggaaact tctggatgct 540 ggtgatttgg atattgaana tgacatgaaa gcacagatgaattgctttta tttgaaancc 600 ttgggatggt tttgttatgg ttctcccnca tgatggtgacatgattttac atttcttgat 660 aatgttgaaa caaatacntt gggattnact tcanttttgaaacttaactg ggaaacantg 720 tgttttgatt tttactccat cccatgtnaa ccat 754<210> SEQ ID NO 13 <211> LENGTH: 3551 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:(29)..(2236) <223> OTHER INFORMATION: CDS <300> PUBLICATION INFORMATION:<308> DATABASE ACCESSION NUMBER: AB073325.1 <309> DATABASE ENTRY DATE:2001-10-23 <313> RELEVANT RESIDUES: (1)..(3551) <400> SEQUENCE: 13gtgaagacat cgcggggacc gattcaccat ggagggcgcc ggcggcgcga acgacaagaa 60aaagataagt tctgaacgtc gaaaagaaaa gtctcgagat gcagccagat ctcggcgaag 120taaagaatct gaagtttttt atgagcttgc tcatcagttg ccacttccac ataatgtgag 180ttcgcatctt gataaggcct ctgtgatgag gcttaccatc agctatttgc gtgtgaggaa 240acttctggat gctggtgatt tggatattga agatgacatg aaagcacaga tgaattgctt 300ttatttgaaa gccttggatg gttttgttat ggttctcaca gatgatggtg acatgattta 360catttctgat aatgtgaaca aatacatggg attaactcag tttgaactaa ctggacacag 420tgtgtttgat tttactcatc catgtgacca tgaggaaatg agagaaatgc ttacacacag 480aaatggcctt gtgaaaaagg gtaaagaaca aaacacacag cgaagctttt ttctcagaat 540gaagtgtacc ctaactagcc gaggaagaac tatgaacata aagtctgcaa catggaaggt 600attgcactgc acaggccaca ttcacgtata tgataccaac agtaaccaac ctcagtgtgg 660gtataagaaa ccacctatga cctgcttggt gctgatttgt gaacccattc ctcacccatc 720aaatattgaa attcctttag atagcaagac tttcctcagt cgacacagcc tggatatgaa 780attttcttat tgtgatgaaa gaattaccga attgatggga tatgagccag aagaactttt 840aggccgctca atttatgaat attatcatgc tttggactct gatcatctga ccaaaactca 900tcatgatatg tttactaaag gacaagtcac cacaggacag tacaggatgc ttgccaaaag 960aggtggatat gtctgggttg aaactcaagc aactgtcata tataacacca agaattctca 1020accacagtgc attgtatgtg tgaattacgt tgtgagtggt attattcagc acgacttgat 1080tttctccctt caacaaacag aatgtgtcct taaaccggtt gaatcttcag atatgaaaat 1140gactcagcta ttcaccaaag ttgaatcaga agatacaagt agcctctttg acaaacttaa 1200gaaggaacct gatgctttaa ctttgctggc cccagccgct ggagacacaa tcatatcttt 1260agattttggc agcaacgaca cagaaactga tgaccagcaa cttgaggaag taccattata 1320taatgatgta atgctcccct cacccaacga aaaattacag aatataaatt tggcaatgtc 1380tccattaccc accgctgaaa cgccaaagcc acttcgaagt agtgctgacc ctgcactcaa 1440tcaagaagtt gcattaaaat tagaaccaaa tccagagtca ctggaacttt cttttaccat 1500gccccagatt caggatcaga cacctagtcc ttccgatgga agcactagac aaagttcacc 1560tgagcctaat agtcccagtg aatattgttt ttatgtggat agtgatatgg tcaatgaatt 1620caagttggaa ttggtagaaa aactttttgc tgaagacaca gaagcaaaga acccattttc 1680tactcaggac acagatttag acttggagat gttagctccc tatatcccaa tggatgatga 1740cttccagtta cgttccttcg atcagttgtc accattagaa agcagttccg caagccctga 1800aagcgcaagt cctcaaagca cagttacagt attccagcag actcaaatac aagaacctac 1860tgctaatgcc accactacca ctgccaccac tgatgaatta aaaacagtga caaaagaccg 1920tatggaagac attaaaatat tgattgcatc tccatctcct acccacatac ataaagaaac 1980tactagtgcc acatcatcac catatagaga tactcaaagt cggacagcct caccaaacag 2040agcaggaaaa ggagtcatag aacagacaga aaaatctcat ccaagaagcc ctaacgtgtt 2100atctgtcgct ttgagtcaaa gaactacagt tcctgaggaa gaactaaatc caaagatact 2160agctttgcag aatgctcaga gaaagcgaaa aatggaacat gatggttcac tttttcaagc 2220agtaggaatt atttagcatg tagactgctg gggcaatcaa tggatgaaag tggattacca 2280cagctgacca gttatgattg tgaagttaat gctcctatac aaggcagcag aaacctactg 2340cagggtgaag aattactcag agctttggat caagttaact gagctttttc ttaatttcat 2400tccttttttt ggacactggt ggctcactac ctaaagcagt ctatttatat tttctacatc 2460taattttaga agcctggcta caatactgca caaacttggt tagttcaatt tttgatcccc 2520tttctactta atttacatta atgctctttt ttagtatgtt ctttaatgct ggatcacaga 2580cagctcattt tctcagtttt ttggtattta aaccattgca ttgcagtagc atcattttaa 2640aaaatgcacc tttttattta tttatttttg gctagggagt ttatcccttt ttcgaattat 2700ttttaagaag atgccaatat aatttttgta agaaggcagt aacctttcat catgatcata 2760ggcagttgaa aaatttttac accttttttt tcacatttta cataaataat aatgctttgc 2820cagcagtacg tggtagccac aattgcacaa tatattttct taaaaaatac cagcagttac 2880tcatggaata tattctgcgt ttataaaact agtttttaag aagaaatttt ttttggccta 2940tgaaattgtt aaacctggaa catgacattg ttaatcatat aataatgatt cttaaatgct 3000gtatggttta ttatttaaat gggtaaagcc atttacataa tatagaaaga tatgcatata 3060tctagaaggt atgtggcatt tatttggata aaattctcaa ttcagagaaa tcatctgatg 3120tttctatagt cactttgcca gctcaaaaga aaacaatacc ctatgtagtt gtggaagttt 3180atgctaatat tgtgtaactg atattaaacc taaatgttct gcctaccctg ttggtataaa 3240gatattttga gcagactgta aacaagaaaa aaaaaatcat gcattcttag caaaattgcc 3300tagtatgtta atttgctcaa aatacaatgt ttgattttat gcactttgtc gctattaaca 3360tccttttttt catgtagatt tcaataattg agtaatttta gaagcattat tttaggaata 3420tatagttgtc acagtaaata tcttgttttt tctatgtaca ttgtacaaat ttttcattcc 3480ttttgctctt tgtggttgga tctaacacta actgtattgt tttgttacat caaataaaca 3540tcttctgtgg a 3551 <210> SEQ ID NO 14 <211> LENGTH: 20 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 14 aaagtgatgtagtagctgca 20 <210> SEQ ID NO 15 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 15 ggtatcatat acgtgaatgt 20 <210>SEQ ID NO 16 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 16 taccacgtac tgctggcaaa 20 <210> SEQ ID NO 17 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:17 tgtgctttga ggacttgcgc 20 <210> SEQ ID NO 18 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 18 gaaatgtaaatcatgtcacc 20 <210> SEQ ID NO 19 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 19 tcaaagaggc tacttgtatc 20 <210>SEQ ID NO 20 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 20 ttaatgcaac ttcttgattg 20 <210> SEQ ID NO 21 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:21 atcattatta tatgattaac 20 <210> SEQ ID NO 22 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 22 gaaaggcaagtccagaggtg 20 <210> SEQ ID NO 23 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 23 taaactccct agccaaaaat 20 <210>SEQ ID NO 24 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 24 cattagcagt aggttcttgt 20 <210> SEQ ID NO 25 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:25 gatcatgatg aaaggttact 20 <210> SEQ ID NO 26 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 26 aaatttcatatccaggctgt 20 <210> SEQ ID NO 27 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 27 agtttcctca cacgcaaata 20 <210>SEQ ID NO 28 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 28 actgatcgaa ggaacgtaac 20 <210> SEQ ID NO 29 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:29 cgctttctct gagcattctg 20 <210> SEQ ID NO 30 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 30 aaatcaaacacactgtgtcc 20 <210> SEQ ID NO 31 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 31 tcctttagta aacatatcat 20 <210>SEQ ID NO 32 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 32 caaagttaaa gcatcaggtt 20 <210> SEQ ID NO 33 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:33 ctagtgcttc catcggaagg 20 <210> SEQ ID NO 34 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 34 aatgccacataccttctaga 20 <210> SEQ ID NO 35 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 35 tcgtgagact agagagaagc 20 <210>SEQ ID NO 36 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 36 atgaaaggtt actgccttct 20 <210> SEQ ID NO 37 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:37 tcagcaccaa gcaggtcata 20 <210> SEQ ID NO 38 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 38 aagtttgtgcagtattgtag 20 <210> SEQ ID NO 39 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 39 ctgagcattc tgcaaagcta 20 <210>SEQ ID NO 40 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 40 ttcagattct ttacttcgcc 20 <210> SEQ ID NO 41 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:41 gataacacgt tagggcttct 20 <210> SEQ ID NO 42 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 42 tcaaagcgacagataacacg 20 <210> SEQ ID NO 43 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 43 caaagcatga taatattcat 20 <210>SEQ ID NO 44 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 44 ccatcatctg tgagaaccat 20 <210> SEQ ID NO 45 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:45 atatggtgat gatgtggcac 20 <210> SEQ ID NO 46 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 46 ctcctcaggtggcttgtcag 20 <210> SEQ ID NO 47 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 47 tgagctgtct gtgatccagc 20 <210>SEQ ID NO 48 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 48 agataacacg ttagggcttc 20 <210> SEQ ID NO 49 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:49 catggtgaat cggtccccgc 20 <210> SEQ ID NO 50 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 50 tgttatatatgacagttgct 20 <210> SEQ ID NO 51 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 51 ccttatcaag atgcgaactc 20 <210>SEQ ID NO 52 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 52 ccaaatcacc agcatccaga 20 <210> SEQ ID NO 53 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:53 aactgagtta atcccatgta 20 <210> SEQ ID NO 54 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 54 ttagttcaaactgagttaat 20 <210> SEQ ID NO 55 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 55 aggccatttc tgtgtgtaag 20 <210>SEQ ID NO 56 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 56 ctatctaaag gaatttcaat 20 <210> SEQ ID NO 57 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:57 cccatcaatt cggtaattct 20 <210> SEQ ID NO 58 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 58 tatcatgatgagttttggtc 20 <210> SEQ ID NO 59 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 59 aataatacca ctcacaacgt 20 <210>SEQ ID NO 60 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 60 caactttggt gaatagctga 20 <210> SEQ ID NO 61 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:61 agtgactctg gatttggttc 20 <210> SEQ ID NO 62 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 62 catctccaagtctaaatctg 20 <210> SEQ ID NO 63 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 63 ctaatggtga caactgatcg 20 <210>SEQ ID NO 64 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 64 cactgttttt aattcatcag 20 <210> SEQ ID NO 65 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:65 ataatgttcc aattcctact 20 <210> SEQ ID NO 66 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 66 agaaaaagctcagttaactt 20 <210> SEQ ID NO 67 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 67 attgtagcca ggcttctaaa 20 <210>SEQ ID NO 68 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 68 atcttcttaa aaataattcg 20 <210> SEQ ID NO 69 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:69 tgtgcaattg tggctaccac 20 <210> SEQ ID NO 70 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 70 aacaatgtcatgttccaggt 20 <210> SEQ ID NO 71 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 71 gctggcaaag tgactataga 20 <210>SEQ ID NO 72 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 72 ttccacagaa gatgtttatt 20 <210> SEQ ID NO 73 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:73 tttttccaca gaagatgttt 20 <210> SEQ ID NO 74 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 74 tagagctaaacgatctagaa 20 <210> SEQ ID NO 75 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 75 taactctttc tggccttgaa 20 <210>SEQ ID NO 76 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 76 attggcccta acagaaaatc 20 <210> SEQ ID NO 77 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:77 agaacttatc ctacttaaca 20 <210> SEQ ID NO 78 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 78 gtttccctcgtgttgctcag 20 <210> SEQ ID NO 79 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 79 ttgtacttac tatcatgatg 20 <210>SEQ ID NO 80 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 80 acttacttac ctcacaacgt 20 <210> SEQ ID NO 81 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:81 aatctgtgtc ctttaaaaca 20 <210> SEQ ID NO 82 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 82 tgtgcactgaggagctgagg 20 <210> SEQ ID NO 83 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 83 acgttcagaa cttatctttt 20 <210>SEQ ID NO 84 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 84 catgctaaat aattcctact 20 <210> SEQ ID NO 85 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:85 tgcagctact acatcacttt 20 <210> SEQ ID NO 86 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 86 acattcacgtatatgatacc 20 <210> SEQ ID NO 87 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 87 gcgcaagtcc tcaaagcaca 20 <210>SEQ ID NO 88 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 88 ggtgacatga tttacatttc 20 <210> SEQ ID NO 89<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 89 gatacaagta gcctctttga 20 <210> SEQ ID NO 90 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 90caatcaagaa gttgcattaa 20 <210> SEQ ID NO 91 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 91 gttaatcatataataatgat 20 <210> SEQ ID NO 92 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 92 cacctctgga cttgcctttc 20 <210>SEQ ID NO 93 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 93 atttttggct agggagttta 20 <210> SEQ ID NO 94<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 94 acaagaacct actgctaatg 20 <210> SEQ ID NO 95 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 95agtaaccttt catcatgatc 20 <210> SEQ ID NO 96 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 96 acagcctggatatgaaattt 20 <210> SEQ ID NO 97 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 97 gttacgttcc ttcgatcagt 20 <210>SEQ ID NO 98 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homosapiens <400> SEQUENCE: 98 cagaatgctc agagaaagcg 20 <210> SEQ ID NO 99<211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400>SEQUENCE: 99 ggacacagtg tgtttgattt 20 <210> SEQ ID NO 100 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 100atgatatgtt tactaaagga 20 <210> SEQ ID NO 101 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 101 aacctgatgctttaactttg 20 <210> SEQ ID NO 102 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 102 gcttctctct agtctcacga 20<210> SEQ ID NO 103 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 103 agaaggcagt aacctttcat 20 <210> SEQ IDNO 104 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 104 ggcgaagtaa agaatctgaa 20 <210> SEQ ID NO 105 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:105 agaagcccta acgtgttatc 20 <210> SEQ ID NO 106 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 106 cgtgttatctgtcgctttga 20 <210> SEQ ID NO 107 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 107 atgaatatta tcatgctttg 20<210> SEQ ID NO 108 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 108 atggttctca cagatgatgg 20 <210> SEQ IDNO 109 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 109 gtgccacatc atcaccatat 20 <210> SEQ ID NO 110 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:110 gctggatcac agacagctca 20 <210> SEQ ID NO 111 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 111 gaagccctaacgtgttatct 20 <210> SEQ ID NO 112 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 112 gcggggaccg attcaccatg 20<210> SEQ ID NO 113 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 113 agcaactgtc atatataaca 20 <210> SEQ IDNO 114 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 114 gagttcgcat cttgataagg 20 <210> SEQ ID NO 115 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:115 tacatgggat taactcagtt 20 <210> SEQ ID NO 116 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 116 cttacacacagaaatggcct 20 <210> SEQ ID NO 117 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 117 agaattaccg aattgatggg 20<210> SEQ ID NO 118 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 118 gaccaaaact catcatgata 20 <210> SEQ IDNO 119 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 119 acgttgtgag tggtattatt 20 <210> SEQ ID NO 120 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:120 tcagctattc accaaagttg 20 <210> SEQ ID NO 121 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 121 gaaccaaatccagagtcact 20 <210> SEQ ID NO 122 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 122 cgatcagttg tcaccattag 20<210> SEQ ID NO 123 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 123 ctgatgaatt aaaaacagtg 20 <210> SEQ IDNO 124 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 124 aagttaactg agctttttct 20 <210> SEQ ID NO 125 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:125 tttagaagcc tggctacaat 20 <210> SEQ ID NO 126 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 126 gtggtagccacaattgcaca 20 <210> SEQ ID NO 127 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 127 acctggaaca tgacattgtt 20<210> SEQ ID NO 128 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 128 tctatagtca ctttgccagc 20 <210> SEQ IDNO 129 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 129 ttctagatcg tttagctcta 20 <210> SEQ ID NO 130 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:130 ttcaaggcca gaaagagtta 20 <210> SEQ ID NO 131 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 131 ctgagcaacacgagggaaac 20 <210> SEQ ID NO 132 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 132 aaaagataag ttctgaacgt 20<210> SEQ ID NO 133 <211> LENGTH: 3933 <212> TYPE: DNA <213> ORGANISM:Homo Sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:(265)..(2745) <223> OTHER INFORMATION: CDS <300> PUBLICATIONINFORMATION: <301> AUTHORS: Hogenesch, et al. <302> TITLE:Characterization Of A Subset Of The Basic-Helix-Loop-Helix- PASSuperfamily That Interacts With Components Of The Dioxin Signaling <303>JOURNAL: J. Biol. Chem. <304> VOLUME: 272 <305> ISSUE: 13 <306> PAGES:8581-8593 <307> DATE: 1997 <308> DATABASE ACCESSION NUMBER: U29165.1<309> DATABASE ENTRY DATE: 1997-04-11 <313> RELEVANT RESIDUES:(1)..(3933) <400> SEQUENCE: 133 cacgaggcag cactctcttc gtcgcttcggccagtgtgtc gggctgggcc ctgacaagcc 60 acctgaggag aggctcggag ccgggcccggaccccggcga ttgccgcccg cttctctcta 120 gtctcacgag gggtttcccg cctcgcacccccacctctgg acttgccttt ccttctcttc 180 tccgcgtgtg gagggagcca gcgcttaggccggagcgagc ctgggggccg cccgccgtga 240 agacatcgcg gggaccgatt caccatggagggcgccggcg gcgcgaacga caagaaaaag 300 ataagttctg aacgtcgaaa agaaaagtctcgagatgcag ccagatctcg gcgaagtaaa 360 gaatctgaag ttttttatga gcttgctcatcagttgccac ttccacataa tgtgagttcg 420 catcttgata aggcctctgt gatgaggcttaccatcagct atttgcgtgt gaggaaactt 480 ctggatgctg gtgatttgga tattgaagatgacatgaaag cacagatgaa ttgcttttat 540 ttgaaagcct tggatggttt tgttatggttctcacagatg atggtgacat gatttacatt 600 tctgataatg tgaacaaata catgggattaactcagtttg aactaactgg acacagtgtg 660 tttgatttta ctcatccatg tgaccatgaggaaatgagag aaatgcttac acacagaaat 720 ggccttgtga aaaagggtaa agaacaaaacacacagcgaa gcttttttct cagaatgaag 780 tgtaccctaa ctagccgagg aagaactatgaacataaagt ctgcaacatg gaaggtattg 840 cactgcacag gccacattca cgtatatgataccaacagta accaacctca gtgtgggtat 900 aagaaaccac ctatgacctg cttggtgctgatttgtgaac ccattcctca cccatcaaat 960 attgaaattc ctttagatag caagactttcctcagtcgac acagcctgga tatgaaattt 1020 tcttattgtg atgaaagaat taccgaattgatgggatatg agccagaaga acttttaggc 1080 cgctcaattt atgaatatta tcatgctttggactctgatc atctgaccaa aactcatcat 1140 gatatgttta ctaaaggaca agtcaccacaggacagtaca ggatgcttgc caaaagaggt 1200 ggatatgtct gggttgaaac tcaagcaactgtcatatata acaccaagaa ttctcaacca 1260 cagtgcattg tatgtgtgaa ttacgttgtgagtggtatta ttcagcacga cttgattttc 1320 tcccttcaac aaacagaatg tgtccttaaaccggttgaat cttcagatat gaaaatgact 1380 cagctattca ccaaagttga atcagaagatacaagtagcc tctttgacaa acttaagaag 1440 gaacctgatg ctttaacttt gctggccccagccgctggag acacaatcat atctttagat 1500 tttggcagca acgacacaga aactgatgaccagcaacttg aggaagtacc attatataat 1560 gatgtaatgc tcccctcacc caacgaaaaattacagaata taaatttggc aatgtctcca 1620 ttacccaccg ctgaaacgcc aaagccacttcgaagtagtg ctgaccctgc actcaatcaa 1680 gaagttgcat taaaattaga accaaatccagagtcactgg aactttcttt taccatgccc 1740 cagattcagg atcagacacc tagtccttccgatggaagca ctagacaaag ttcacctgag 1800 cctaatagtc ccagtgaata ttgtttttatgtggatagtg atatggtcaa tgaattcaag 1860 ttggaattgg tagaaaaact ttttgctgaagacacagaag caaagaaccc attttctact 1920 caggacacag atttagactt ggagatgttagctccctata tcccaatgga tgatgacttc 1980 cagttacgtt ccttcgatca gttgtcaccattagaaagca gttccgcaag ccctgaaagc 2040 gcaagtcctc aaagcacagt tacagtattccagcagactc aaatacaaga acctactgct 2100 aatgccacca ctaccactgc caccactgatgaattaaaaa cagtgacaaa agaccgtatg 2160 gaagacatta aaatattgat tgcatctccatctcctaccc acatacataa agaaactact 2220 agtgccacat catcaccata tagagatactcaaagtcgga cagcctcacc aaacagagca 2280 ggaaaaggag tcatagaaca gacagaaaaatctcatccaa gaagccctaa cgtgttatct 2340 gtcgctttga gtcaaagaac tacagttcctgaggaagaac taaatccaaa gatactagct 2400 ttgcagaatg ctcagagaaa gcgaaaaatggaacatgatg gttcactttt tcaagcagta 2460 ggaattggaa cattattaca gcagccagacgatcatgcag ctactacatc actttcttgg 2520 aaacgtgtaa aaggatgcaa atctagtgaacagaatggaa tggagcaaaa gacaattatt 2580 ttaataccct ctgatttagc atgtagactgctggggcaat caatggatga aagtggatta 2640 ccacagctga ccagttatga ttgtgaagttaatgctccta tacaaggcag cagaaaccta 2700 ctgcagggtg aagaattact cagagctttggatcaagtta actgagcttt ttcttaattt 2760 cattcctttt tttggacact ggtggctcactacctaaagc agtctattta tattttctac 2820 atctaatttt agaagcctgg ctacaatactgcacaaactt ggttagttca atttttgatc 2880 ccctttctac ttaatttaca ttaatgctcttttttagtat gttctttaat gctggatcac 2940 agacagctca ttttctcagt tttttggtatttaaaccatt gcattgcagt agcatcattt 3000 taaaaaatgc acctttttat ttatttatttttggctaggg agtttatccc tttttcgaat 3060 tatttttaag aagatgccaa tataatttttgtaagaaggc agtaaccttt catcatgatc 3120 ataggcagtt gaaaaatttt tacaccttttttttcacatt ttacataaat aataatgctt 3180 tgccagcagt acgtggtagc cacaattgcacaatatattt tcttaaaaaa taccagcagt 3240 tactcatgga atatattctg cgtttataaaactagttttt aagaagaaat tttttttggc 3300 ctatgaaatt gttaaacctg gaacatgacattgttaatca tataataatg attcttaaat 3360 gctgtatggt ttattattta aatgggtaaagccatttaca taatatagaa agatatgcat 3420 atatctagaa ggtatgtggc atttatttggataaaattct caattcagag aaatcatctg 3480 atgtttctat agtcactttg ccagctcaaaagaaaacaat accctatgta gttgtggaag 3540 tttatgctaa tattgtgtaa ctgatattaaacctaaatgt tctgcctacc ctgttggtat 3600 aaagatattt tgagcagact gtaaacaagaaaaaaaaaat catgcattct tagcaaaatt 3660 gcctagtatg ttaatttgct caaaatacaatgtttgattt tatgcacttt gtcgctatta 3720 acatcctttt tttcatgtag atttcaataattgagtaatt ttagaagcat tattttagga 3780 atatatagtt gtcacagtaa atatcttgttttttctatgt acattgtaca aatttttcat 3840 tccttttgct ctttgtggtt ggatctaacactaactgtat tgttttgtta catcaaataa 3900 acatcttctg tggaaaaaaa aaaaaaaaaaaaa 3933 <210> SEQ ID NO 134 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 134 tgatgagcaa gctcataaaa 20 <210>SEQ ID NO 135 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 135 gcaactgatg agcaagctca 20 <210> SEQ ID NO136 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 136 ggaagtggca actgatgagc 20 <210> SEQ ID NO 137 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 137ccagttagtt caaactgagt 20 <210> SEQ ID NO 138 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 138 tgtgtccagttagttcaaac 20 <210> SEQ ID NO 139 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 139 cacactgtgt ccagttagtt 20 <210>SEQ ID NO 140 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 140 cacatggatg agtaaaatca 20 <210> SEQ ID NO141 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 141 tcctcatggt cacatggatg 20 <210> SEQ ID NO 142 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 142tctctcattt cctcatggtc 20 <210> SEQ ID NO 143 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 143 gcatttctctcatttcctca 20 <210> SEQ ID NO 144 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 144 gtgtgtaagc atttctctca 20 <210>SEQ ID NO 145 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 145 ggccatttct gtgtgtaagc 20 <210> SEQ ID NO146 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 146 tggttactgt tggtatcata 20 <210> SEQ ID NO 147 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 147tcacaaatca gcaccaagca 20 <210> SEQ ID NO 148 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 148 tgggttcacaaatcagcacc 20 <210> SEQ ID NO 149 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 149 tgaggaatgg gttcacaaat 20 <210>SEQ ID NO 150 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 150 gtcttgctat ctaaaggaat 20 <210> SEQ ID NO151 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 151 tattcataaa ttgagcggcc 20 <210> SEQ ID NO 152 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 152tgataatatt cataaattga 20 <210> SEQ ID NO 153 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 153 tgagttttggtcagatgatc 20 <210> SEQ ID NO 154 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 154 acttgtcctt tagtaaacat 20 <210>SEQ ID NO 155 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 155 tggtgacttg tcctttagta 20 <210> SEQ ID NO156 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 156 tcctgtggtg acttgtcctt 20 <210> SEQ ID NO 157 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 157tactgtcctg tggtgacttg 20 <210> SEQ ID NO 158 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 158 tcctgtactgtcctgtggtg 20 <210> SEQ ID NO 159 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 159 gcaagcatcc tgtactgtcc 20 <210>SEQ ID NO 160 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 160 cagacatatc cacctctttt 20 <210> SEQ ID NO161 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 161 tcaacccaga catatccacc 20 <210> SEQ ID NO 162 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 162tatgacagtt gcttgagttt 20 <210> SEQ ID NO 163 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 163 ttatatatgacagttgcttg 20 <210> SEQ ID NO 164 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 164 gaagggagaa aatcaagtcg 20 <210>SEQ ID NO 165 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 165 attctgtttg ttgaagggag 20 <210> SEQ ID NO166 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 166 ttcatatctg aagattcaac 20 <210> SEQ ID NO 167 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 167tctgattcaa ctttggtgaa 20 <210> SEQ ID NO 168 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 168 attacatcattatataatgg 20 <210> SEQ ID NO 169 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 169 ctacttcgaa gtggctttgg 20 <210>SEQ ID NO 170 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 170 tcagcactac ttcgaagtgg 20 <210> SEQ ID NO171 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 171 ctttgtctag tgcttccatc 20 <210> SEQ ID NO 172 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 172atcatccatt gggatatagg 20 <210> SEQ ID NO 173 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 173 tctaatggtgacaactgatc 20 <210> SEQ ID NO 174 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 174 catcatgttc catttttcgc 20 <210>SEQ ID NO 175 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 175 gtcagctgtg gtaatccact 20 <210> SEQ ID NO176 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 176 taactggtca gctgtggtaa 20 <210> SEQ ID NO 177 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 177ggagcattaa cttcacaatc 20 <210> SEQ ID NO 178 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 178 aggtttctgctgccttgtat 20 <210> SEQ ID NO 179 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 179 ccctgcagta ggtttctgct 20 <210>SEQ ID NO 180 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 180 cttcaccctg cagtaggttt 20 <210> SEQ ID NO181 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 181 taattcttca ccctgcagta 20 <210> SEQ ID NO 182 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 182ctgagtaatt cttcaccctg 20 <210> SEQ ID NO 183 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 183 aagctctgagtaattcttca 20 <210> SEQ ID NO 184 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 184 atccaaagct ctgagtaatt 20 <210>SEQ ID NO 185 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 185 acttgatcca aagctctgag 20 <210> SEQ ID NO186 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 186 gctcagttaa cttgatccaa 20 <210> SEQ ID NO 187 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 187tgagccacca gtgtccaaaa 20 <210> SEQ ID NO 188 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 188 ccaggcttctaaaattagat 20 <210> SEQ ID NO 189 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 189 gtgcagtatt gtagccaggc 20 <210>SEQ ID NO 190 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 190 agtttgtgca gtattgtagc 20 <210> SEQ ID NO191 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 191 taaataaaaa ggtgcatttt 20 <210> SEQ ID NO 192 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 192actgcctatg atcatgatga 20 <210> SEQ ID NO 193 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 193 ttgtgcaattgtggctacca 20 <210> SEQ ID NO 194 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 194 atatattgtg caattgtggc 20 <210>SEQ ID NO 195 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 195 agaaaatata ttgtgcaatt 20 <210> SEQ ID NO196 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 196 cttaaaaact agttttataa 20 <210> SEQ ID NO 197 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 197atgtaaatgg ctttacccat 20 <210> SEQ ID NO 198 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 198 ttttatccaaataaatgcca 20 <210> SEQ ID NO 199 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 199 tgagaatttt atccaaataa 20 <210>SEQ ID NO 200 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 200 taatagcgac aaagtgcata 20 <210> SEQ ID NO201 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 201 gatgttaata gcgacaaagt 20 <210> SEQ ID NO 202 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 202aaaaggatgt taatagcgac 20 <210> SEQ ID NO 203 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 203 aatgcttctaaaattactca 20 <210> SEQ ID NO 204 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 204 tatattccta aaataatgct 20 <210>SEQ ID NO 205 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 205 acagaagatg tttatttgat 20 <210> SEQ ID NO206 <211> LENGTH: 3973 <212> TYPE: DNA <213> ORGANISM: Mus muculus <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (258)..(2768)<223> OTHER INFORMATION: CDS <300> PUBLICATION INFORMATION: <308>DATABASE ACCESSION NUMBER: NM_010431.1 <309> DATABASE ENTRY DATE:2003-12-20 <313> RELEVANT RESIDUES: (1)..(3973) <400> SEQUENCE: 206cgcgaggact gtcctcgccg ccgtcgcggg cagtgtctag ccaggccttg acaagctagc 60cggaggagcg cctaggaacc cgagccggag ctcagcgagc gcagcctgca cgcccgcctc 120gcgtcccggg ggggtcccgc ctcccacccc gcctctggac ttgtctcttt ccccgcgcgc 180gcggacagag ccggcgttta ggcccgagcg agcccggggg ccgccggccg ggaagacaac 240gcgggcaccg attcgccatg gagggcgccg gcggcgagaa cgagaagaaa aagatgagtt 300ctgaacgtcg aaaagaaaag tctagagatg cagcaagatc tcggcgaagc aaagagtctg 360aagtttttta tgagcttgct catcagttgc cacttcccca caatgtgagc tcacatcttg 420ataaagcttc tgttatgagg ctcaccatca gttatttacg tgtgagaaaa cttctggatg 480ccggtggtct agacagtgaa gatgagatga aggcacagat ggactgtttt tatctgaaag 540ccctagatgg ctttgtgatg gtgctaacag atgacggcga catggtttac atttctgata 600acgtgaacaa atacatgggg ttaactcagt ttgaactaac tggacacagt gtgtttgatt 660ttactcatcc atgtgaccat gaggaaatga gagaaatgct tacacacaga aatggcccag 720tgagaaaagg gaaagaacta aacacacagc ggagcttttt tctcagaatg aagtgcaccc 780taacaagccg ggggaggacg atgaacatca agtcagcaac gtggaaggtg cttcactgca 840cgggccatat tcatgtctat gataccaaca gtaaccaacc tcagtgtggg tacaagaaac 900cacccatgac gtgcttggtg ctgatttgtg aacccattcc tcatccgtca aatattgaaa 960ttcctttaga tagcaagaca tttctcagtc gacacagcct cgatatgaaa ttttcttact 1020gtgatgaaag aattactgag ttgatgggtt atgagccgga agaacttttg ggccgctcaa 1080tttatgaata ttatcatgct ttggattctg atcatctgac caaaactcac catgatatgt 1140ttactaaagg acaagtcacc acaggacagt acaggatgct tgccaaaaga ggtggatatg 1200tctgggttga aactcaagca actgtcatat ataatacgaa gaactcccag ccacagtgca 1260ttgtgtgtgt gaattatgtt gtaagtggta ttattcagca cgacttgatt ttctcccttc 1320aacaaacaga atctgtgctc aaaccagttg aatcttcaga tatgaagatg actcagctgt 1380tcaccaaagt tgaatcagag gatacaagct gcctttttga taagcttaag aaggagcctg 1440atgctctcac tctgctggct ccagctgccg gcgacaccat catctctctg gattttggca 1500gcgatgacac agaaactgaa gatcaacaac ttgaagatgt tccattatat aatgatgtaa 1560tgtttccctc ttctaatgaa aaattaaata taaacctggc aatgtctcct ttaccttcat 1620cggaaactcc aaagccactt cgaagtagtg ctgatcctgc actgaatcaa gaggttgcat 1680taaaattaga atcaagtcca gagtcactgg gactttcttt taccatgccc cagattcaag 1740atcagccagc aagtccttct gatggaagca ctagacaaag ttcacctgag agacttcttc 1800aggaaaacgt aaacactcct aacttttccc agcctaacag tcccagtgaa tattgctttg 1860atgtggatag cgatatggtc aatgtattca agttggaact ggtggaaaaa ctgtttgctg 1920aagacacaga ggcaaagaat ccattttcaa ctcaggacac tgatttagat ttggagatgc 1980tggctcccta tatcccaatg gatgatgatt tccagttacg ttcctttgat cagttgtcac 2040cattagagag caattctcca agccctccaa gtatgagcac agttactggg ttccagcaga 2100cccagttaca gaaacctacc atcactgcca ctgccaccac aactgccacc actgatgaat 2160caaaaacaga gacgaaggac aataaagaag atattaaaat actgattgca tctccatctt 2220ctacccaagt acctcaagaa acgaccactg ctaaggcatc agcatacagt ggcactcaca 2280gtcggacagc ctcaccagac agagcaggaa agagagtcat agaacagaca gacaaagctc 2340atccaaggag ccttaagctg tctgccactt tgaatcaaag aaatactgtt cctgaggaag 2400aattaaaccc aaagacaata gcttcgcaga atgctcagag gaagcgaaaa atggaacatg 2460atggctccct ttttcaagca gcaggaattg gaacattatt gcagcaacca ggtgactgtg 2520cacctactat gtcactttcc tggaaacgag tgaaaggatt catatctagt gaacagaatg 2580gaacggagca aaagactatt attttaatac cctccgattt agcatgcaga ctgctggggc 2640agtcaatgga tgagagtgga ttaccacagc tgaccagtta cgattgtgaa gttaatgctc 2700ccatacaagg cagcagaaac ctactgcagg gtgaagaatt actcagagct ttggatcaag 2760ttaactgagc gtttcctaat ctcattcctt ttgattgtta atgtttttgt tcagttgttg 2820ttgtttgttg ggtttttgtt tctgttggtt atttttggac actggtggct cagcagtcta 2880tttatatttt ctatatctaa ttttagaagc ctggctacaa tactgcacaa actcagatag 2940tttagttttc atcccctttc tacttaattt tcattaatgc tctttttaat atgttctttt 3000aatgccagat cacagcacat tcacagctcc tcagcatttc accattgcat tgctgtagtg 3060tcatttaaaa tgcacctttt tatttattta tttttggtga gggagtttgt cccttattga 3120attattttta atgaaatgcc aatataattt tttaagaaag cagtaaattc tcatcatgat 3180cataggcagt tgaaaacttt ttactcattt ttttcatgtt ttacatgaaa ataatgcttt 3240gtcagcagta catggtagcc acaattgcac aatatatttt ctttaaaaaa ccagcagtta 3300ctcatgcaat atattctgca tttataaaac tagtttttaa gaaatttttt ttggcctatg 3360gaattgttaa gcctggatca tgaagcgttg atcttataat gattcttaaa ctgtatggtt 3420tctttatatg ggtaaagcca tttacatgat ataaagaaat atgcttatat ctggaaggta 3480tgtggcattt atttggataa aattctcaat tcagagaagt tatctggtgt ttcttgactt 3540taccaactca aaacagtccc tctgtagttg tggaagctta tgctaatatt gtgtaattga 3600ttatgaaaca taaatgttct gcccaccctg ttggtataaa gacattttga gcatactgta 3660aacaaacaaa caaaaaatca tgctttgtta gtaaaattgc ctagtatgtt gatttgttga 3720aaatatgatg tttggtttta tgcactttgt cgctattaac atcctttttt catatagatt 3780tcaataagtg agtaatttta gaagcattat tttaggaata tagagttgtc atagtaaaca 3840tcttgttttt tctatgtaca ctgtataaat ttttcgttcc cttgctcttt gtggttgggt 3900ctaacactaa ctgtactgtt ttgttatatc aaataaacat cttctgtgga ccaggaaaaa 3960aaaaaaaaaa aaa 3973 <210> SEQ ID NO 207 <211> LENGTH: 20 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 207 gatcatgatgagaatttact 20 <210> SEQ ID NO 208 <211> LENGTH: 2818 <212> TYPE: DNA<213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: (150)..(2762) <223> OTHER INFORMATION: CDS <300>PUBLICATION INFORMATION: <301> AUTHORS: Sang et al. <302> TITLE: MAPKSignaling Up-Regulates The Activity Of Hypoxia- Inducible Factors By ItsEffect on P300 <303> JOURNAL: J. Biol. Chem. <304> VOLUME: 278 <305>ISSUE: 16 <306> PAGES: 14013-14019 <307> DATE: 2003 <308> DATABASEACCESSION NUMBER: NM_001430.1 <309> DATABASE ENTRY DATE: 2003-10-06<313> RELEVANT RESIDUES: (1)..(2818) <400> SEQUENCE: 208 cctgactgcgcggggcgctc gggacctgcg cgcacctcgg accttcacca cccgcccggg 60 ccgcggggagcggacgaggg ccacagcccc ccacccgcca gggagcccag gtgctcggcg 120 tctgaacgtctcaaagggcc acagcgacaa tgacagctga caaggagaag aaaaggagta 180 gctcggagaggaggaaggag aagtcccggg atgctgcgcg gtgccggcgg agcaaggaga 240 cggaggtgttctatgagctg gcccatgagc tgcctctgcc ccacagtgtg agctcccatc 300 tggacaaggcctccatcatg cgactggaaa tcagcttcct gcgaacacac aagctcctct 360 cctcagtttgctctgaaaac gagtccgaag ccgaagctga ccagcagatg gacaacttgt 420 acctgaaagccttggagggt ttcattgccg tggtgaccca agatggcgac atgatctttc 480 tgtcagaaaacatcagcaag ttcatgggac ttacacaggt ggagctaaca ggacatagta 540 tctttgacttcactcatccc tgcgaccatg aggagattcg tgagaacctg agtctcaaaa 600 atggctctggttttgggaaa aaaagcaaag acatgtccac agagcgggac ttcttcatga 660 ggatgaagtgcacggtcacc aacagaggcc gtactgtcaa cctcaagtca gccacctgga 720 aggtcttgcactgcacgggc caggtgaaag tctacaacaa ctgccctcct cacaatagtc 780 tgtgtggctacaaggagccc ctgctgtcct gcctcatcat catgtgtgaa ccaatccagc 840 acccatcccacatggacatc cccctggata gcaagacctt cctgagccgc cacagcatgg 900 acatgaagttcacctactgt gatgacagaa tcacagaact gattggttac caccctgagg 960 agctgcttggccgctcagcc tatgaattct accatgcgct agactccgag aacatgacca 1020 agagtcaccagaacttgtgc accaagggtc aggtagtaag tggccagtac cggatgctcg 1080 caaagcatgggggctacgtg tggctggaga cccaggggac ggtcatctac aaccctcgca 1140 acctgcagccccagtgcatc atgtgtgtca actacgtcct gagtgagatt gagaagaatg 1200 acgtggtgttctccatggac cagactgaat ccctgttcaa gccccacctg atggccatga 1260 acagcatctttgatagcagt ggcaaggggg ctgtgtctga gaagagtaac ttcctattca 1320 ccaagctaaaggaggagccc gaggagctgg cccagctggc tcccacccca ggagacgcca 1380 tcatctctctggatttcggg aatcagaact tcgaggagtc ctcagcctat ggcaaggcca 1440 tcctgcccccgagccagcca tgggccacgg agttgaggag ccacagcacc cagagcgagg 1500 ctgggagcctgcctgccttc accgtgcccc aggcagctgc cccgggcagc accaccccca 1560 gtgccaccagcagcagcagc agctgctcca cgcccaatag ccctgaagac tattacacat 1620 ctttggataacgacctgaag attgaagtga ttgagaagct cttcgccatg gacacagagg 1680 ccaaggaccaatgcagtacc cagacggatt tcaatgagct ggacttggag acactggcac 1740 cctatatccccatggacggg gaagacttcc agctaagccc catctgcccc gaggagcggc 1800 tcttggcggagaacccacag tccacccccc agcactgctt cagtgccatg acaaacatct 1860 tccagccactggcccctgta gccccgcaca gtcccttcct cctggacaag tttcagcagc 1920 agctggagagcaagaagaca gagcccgagc accggcccat gtcctccatc ttctttgatg 1980 ccggaagcaaagcatccctg ccaccgtgct gtggccaggc cagcacccct ctctcttcca 2040 tggggggcagatccaatacc cagtggcccc cagatccacc attacatttt gggcccacaa 2100 agtgggccgtcggggatcag cgcacagagt tcttgggagc agcgccgttg gggccccctg 2160 tctctccaccccatgtctcc accttcaaga caaggtctgc aaagggtttt ggggctcgag 2220 gcccagacgtgctgagtccg gccatggtag ccctctccaa caagctgaag ctgaagcgac 2280 agctggagtatgaagagcaa gccttccagg acctgagcgg gggggaccca cctggtggca 2340 gcacctcacatttgatgtgg aaacggatga agaacctcag gggtgggagc tgccctttga 2400 tgccggacaagccactgagc gcaaatgtac ccaatgataa gttcacccaa aaccccatga 2460 ggggcctgggccatcccctg agacatctgc cgctgccaca gcctccatct gccatcagtc 2520 ccggggagaacagcaagagc aggttccccc cacagtgcta cgccacccag taccaggact 2580 acagcctgtcgtcagcccac aaggtgtcag gcatggcaag ccggctgctc gggccctcat 2640 ttgagtcctacctgctgccc gaactgacca gatatgactg tgaggtgaac gtgcccgtgc 2700 tgggaagctccacgctcctg caaggagggg acctcctcag agccctggac caggccacct 2760 gagccaggccttctacctgg gcagcacctc tgccgacgcc gtcccaccag cttcaccc 2818 <210> SEQ IDNO 209 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer<400> SEQUENCE: 209 aagccttgga gggtttcatt g 21 <210> SEQ ID NO 210 <211>LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE:210 tgctgatgtt ttctgacaga aagat 25 <210> SEQ ID NO 211 <211> LENGTH: 23<212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223>OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE: 211 cgtggtgacccaagatggcg aca 23 <210> SEQ ID NO 212 <211> LENGTH: 3415 <212> TYPE: DNA<213> ORGANISM: Mus musculus <220> FEATURE: <221> NAME/KEY: misc_feature<222> LOCATION: (184)..(2808) <223> OTHER INFORMATION: CDS <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (2965)..(2965)<223> OTHER INFORMATION: n is a, c, g, or t <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: (2969)..(2969) <223> OTHERINFORMATION: n is a, c, g, or t <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (3022)..(3022) <223> OTHER INFORMATION: nis a, c, g, or t <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (3075)..(3075) <223> OTHER INFORMATION: n is a, c, g, or t<300> PUBLICATION INFORMATION: <301> AUTHORS: Compernolle et al. <302>TITLE: Loss of HIF-2 Alpha And Inhibition of VEGF Impair Fetal LungMaturation, Whereas Treatment With VEGF Prevents Fatal RespiratoryDistress In Premature Mice <303> JOURNAL: Nat. Med. <304> VOLUME: 8<305> ISSUE: 7 <306> PAGES: 702-710 <307> DATE: 2002 <308> DATABASEACCESSION NUMBER: NM_010137.1 <309> DATABASE ENTRY DATE: 2003-04-07<313> RELEVANT RESIDUES: (1)..(3415) <400> SEQUENCE: 212 ctagccagccctctgcaaac ttccacctga ttgagcggga ctctcggacc tgcgagcact 60 aaagacctttcacacctgcc cgggcgacag agagctgcgg agggccacag caaagagagc 120 ggctgcagcccctacggggt taaggaaccc aggtgctccg ggtctcggag ggccacggcg 180 acaatgacagctgacaagga gaaaaaaagg agcagctcag agctgaggaa ggagaaatcc 240 cgtgatgccgcgaggtgccg gcgcagcaag gagacggagg tcttctatga gttggctcat 300 gagttgcccctgcctcacag tgtgagctcc cacctggaca aagcctccat catgcgcctg 360 gccatcagcttccttcggac acataagctc ctgtcctcag tctgctctga aaatgaatct 420 gaagctgaggccgaccagca aatggataac ttgtacctga aagccttgga gggtttcatt 480 gctgtggtgacccaagacgg tgacatgatc tttctgtcgg aaaacatcag caagttcatg 540 ggacttacccaggtagaact aacaggacac agcatctttg acttcactca tccttgcgac 600 catgaggagatccgtgagaa cctgactctc aaaaacggct ctggttttgg gaagaagagc 660 aaagacgtgtccaccgagcg tgacttcttc atgaggatga agtgcacagt caccaacaga 720 ggccggactgtcaacctcaa gtcggccacc tggaaggtcc tgcactgcac cgggcaagtg 780 agagtctacaacaactgccc ccctcacagt agtctctgtg gctccaagga gcccctgctg 840 tcctgccttatcatcatgtg tgagccaatc cagcacccat cccacatgga catccccctg 900 gacagcaagactttcctgag ccgccacagc atggacatga agttcaccta ctgtgacgac 960 agaatcttggaactgattgg ttaccacccc gaggagctac ttggacgctc tgcctatgag 1020 ttctaccatgccctggattc ggagaacatg accaaaagtc accagaactt gtgcaccaag 1080 gggcaggtggtatctggcca gtaccggatg ctagccaaac acggaggata tgtgtggctg 1140 gagacccaggggacggtcat ctacaacccc cgcaacctgc agcctcagtg tatcatgtgt 1200 gtcaactatgtgctgagtga gatcgagaag aacgacgtgg tgttctccat ggaccagacc 1260 gaatccctgttcaagccaca cctgatggcc atgaacagca tctttgacag cagtgacgat 1320 gtggctgtaactgagaagag caactacctg ttcaccaaac tgaaggagga gcccgaggag 1380 ctggcccagttggcccccac cccaggagat gccattattt ctctcgattt cggaagccag 1440 aacttcgatgaaccctcagc ctatggcaag gccatccttc ccccgggcca gccatgggcc 1500 gcggggctgaggagccacag tgcccagagc gagtccggga gcctgccagc cttcactgtg 1560 ccccaggcaggcaccccagg gaacactaca cccagtgctt caagcagcag tagctgctcc 1620 acgcccagcagccctgagga ctactattca tccttggaga atcccttgaa gatcgaagtg 1680 attgagaagcttttcgccat ggacacggag ccgagggacc cgggcagtac ccagacggac 1740 ttcagtgaactggatttgga gaccttggca ccctacatcc ctatggacgg cgaggacttc 1800 cagctgagccccatctgccc agaggagccg ctcatgccag agagccccca gcccaccccc 1860 cagcactgcttcagtaccat gaccagcatc ttccagccgc tcaccccggg ggccacccac 1920 ggccccttcttcctcgataa gtacccgcag cagttggaaa gcaggaagac agagtctgag 1980 cactggcccatgtcttccat cttctttgat gctgggagca aagggtccct gtctccatgc 2040 tgtggccaggccagcacccc tctctcttct atggggggca gatccaacac gcagtggccc 2100 ccggatccaccattacattt cggccctact aagtggcctg tgggtgatca gagtgctgaa 2160 tccctgggagccctgccggt ggggtcatcg cagttggaac ctccgagcgc cccgcctcat 2220 gtctccatgttcaagatgag gtctgcaaag gacttcgggg cccgaggtcc atacatgatg 2280 agcccagccatgatcgccct gtccaacaag ctgaagctaa agcggcagct ggagtatgag 2340 gagcaagctttccaagaaac aagcgggggg gaccttccag gcaccagcag ttcacacttg 2400 atgtggaaacgtatgaagag cctcatgggc gggacctgtc ctttgatgcc tgacaagacc 2460 atcagtgggaacatggcccc ggatgaattc acccaaaaat ctatgagagg cttgggccag 2520 ccattgagacacttgccact tccccagcca ccatttacca ggaactcagg ggagaacgcc 2580 aagactgggttcccgccaca gtgctatgcc tcccagttcc aggactacgg tcctccagga 2640 gctcaaaaggtgtcaggcgt ggccagtcga ctgctggggc catcgttcga gccttacctg 2700 ttgccggaactgaccagata tgactgtgag gtgaacgtgc ccgtgcctgg aagctccaca 2760 ctcctgcaggggagagacct tctcagagct ctggaccagg ccacctgagc cagggcctct 2820 ggccgggcatgcccctgcct gccccgccgt cttgacctgc cagcttcact tccatctgtg 2880 ttgctattaggtatctctaa caccagcaca cttcttacga gatgtactca acctggccta 2940 ctggccaggtcaccaagcag tggcntttnt ctgacatgct cactttatta tccatgtttt 3000 aaaaatacatagttgttgta cntgctatgt tttaccgttg atgaaagtgt tctgaaattt 3060 tataagatttccccntcctt ccttcccttg aattacttct aatttatatt ccccaaaggt 3120 ttttctctctctcattcata tccatactaa caagcatggt ggctggtgcc tctccctagg 3180 aaagctttggcgtcattcaa ctcaagtgtt cttgttcttg ttgccaaaga gaaaaggatt 3240 ttcctccactgtggattttc cctctccccc acccccacat acacacacac acacacacac 3300 acacccctacacacatatac acacatgcac gtatgcgtgc acacacacac acacacatat 3360 acacacacacacacacacac acacacaccc ctacacacat atacacacat gcacc 3415 <210> SEQ ID NO213 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Oligonucleotide primer <400>SEQUENCE: 213 ggccatcgtt cgagcctta 19 <210> SEQ ID NO 214 <211> LENGTH:16 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE: 214ggcacgggca cgttca 16 <210> SEQ ID NO 215 <211> LENGTH: 29 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 215 ctgttgccggaactgaccag atatgactg 29 <210> SEQ ID NO 216 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 216 ggcaaattcaacggcacagt 20 <210> SEQ ID NO 217 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 217 gggtctcgct cctggaagat 20<210> SEQ ID NO 218 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 218 aaggccgaga atgggaagct tgtcatc27 <210> SEQ ID NO 219 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 219 gtcagctgtc attgtcgctg 20<210> SEQ ID NO 220 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 220 ggcctggctc aggtggcctg 20<210> SEQ ID NO 221 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 221 ggtcatgttc tcggagtcta 20<210> SEQ ID NO 222 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 222 gtggagcagc tgctgctgct 20<210> SEQ ID NO 223 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 223 ggtacatttg cgctcagtgg 20<210> SEQ ID NO 224 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 224 tgggcctcga gccccaaaac 20<210> SEQ ID NO 225 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 225 gaataggaag ttactcttct 20<210> SEQ ID NO 226 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 226 tggaagtctt ccccgtccat 20<210> SEQ ID NO 227 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 227 gcagctcctc agggtggtaa 20<210> SEQ ID NO 228 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 228 catggtagaa ttcataggct 20<210> SEQ ID NO 229 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 229 tcacttcaat cttcaggtcg 20<210> SEQ ID NO 230 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 230 gagcttccca gcacgggcac 20<210> SEQ ID NO 231 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 231 tgaaggcagg caggctccca 20<210> SEQ ID NO 232 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 232 ggtgctggcc tggccacagc 20<210> SEQ ID NO 233 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 233 cgaatctcct catggtcgca 20<210> SEQ ID NO 234 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 234 tgctgttcat ggccatcagg 20<210> SEQ ID NO 235 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 235 tactgcattg gtccttggcc 20<210> SEQ ID NO 236 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 236 ctcccagcct cgctctgggt 20<210> SEQ ID NO 237 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 237 aggagcgtgg agcttcccag 20<210> SEQ ID NO 238 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 238 ctgtggacat gtctttgctt 20<210> SEQ ID NO 239 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 239 agtgtctcca agtccagctc 20<210> SEQ ID NO 240 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 240 ctattgtgag gagggcagtt 20<210> SEQ ID NO 241 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 241 tcatagaaca cctccgtctc 20<210> SEQ ID NO 242 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 242 aaatgtgagg tgctgccacc 20<210> SEQ ID NO 243 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 243 ttgggcgtgg agcagctgct 20<210> SEQ ID NO 244 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 244 gcgctgctcc caagaactct 20<210> SEQ ID NO 245 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 245 gcagcaggta ggactcaaat 20<210> SEQ ID NO 246 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 246 gtgctgccac caggtgggtc 20<210> SEQ ID NO 247 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 247 tggtcatgtt ctcggagtct 20<210> SEQ ID NO 248 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 248 tcagtctggt ccatggagaa 20<210> SEQ ID NO 249 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 249 tctcacgaat ctcctcatgg 20<210> SEQ ID NO 250 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 250 tcttcaggtc gttatccaaa 20<210> SEQ ID NO 251 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 251 aggtcccctc cttgcaggag 20<210> SEQ ID NO 252 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 252 tgggccagct catagaacac 20<210> SEQ ID NO 253 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 253 tcaaatgtga ggtgctgcca 20<210> SEQ ID NO 254 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 254 catctgctgg tcagcttcgg 20<210> SEQ ID NO 255 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 255 acagggattc agtctggtcc 20<210> SEQ ID NO 256 <211> LENGTH: 0 <212> TYPE: DNA <213> ORGANISM: ...<400> SEQUENCE: 256 000 <210> SEQ ID NO 257 <211> LENGTH: 78695 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 257 aacaagtcgcagttctgcgg aaggaggagg aagggcaggc gggcgcccga ggagcgcagc 60 tccagagaaaggggagggac ggggtcaggg cagaggctgt ggccgcgcct ccctattggc 120 cgggacacggtgggaagtcg gggcaggcag ccagcgtgcg gggcggggct tctctggagc 180 cccgcccccggcccgccccc agcccgatgt cccactcccg cgcccctcgc gccctcccgc 240 gggcgggccgtccgagtttt taaagtgggc tgacagccga ggaggccgga cattcgagag 300 cggccggtgtacagctccgg agtccgcagc gctccgctcc agctctcctg aggcggccgt 360 acaatcctcggcagtgtcct gagactgtat ggtcatctca gcggccgcac tcgcttgccc 420 ccggatttttttccaacttg ctctcttcga gccatttttt tttctttttt tctttttctt 480 tttttctttttcttttttgg ttggttggtt ttgatttgtc agatcccaga aaagtgactc 540 ctgttcggggctaaacggaa ctccaggtcc cttgtcgctg ctctctctct cttttggcgt 600 cttacaacctcctcccactc ctttccccgg ccccgcctcc tcctgcaggt tcctccccgt 660 cacccccctcctccctcctc ctcctccgca cctagccagc cctctgcaaa cttccacctg 720 attgagcgggactctcggac ctgcgagcac taaagacctt tcacacctgc ccgggcgaca 780 gagagctgcggagggccaca gcaaagagag cggctgcagc ccctacgggg ttaaggaacc 840 caggtgctccgggtctcgga gggccacggc gacaatgaca gctgacaagg agaaaaaaag 900 gtaagcgggagtcaaagctg gcgaccgact tggtcccagg cattggcgga actgcgcgtg 960 gcgggcgcgacccggggcag tcgggagagg ggtgcgagag ttctcggagg ctggaggacc 1020 gctgttgaggggctttgagt aagcgagtcg gtgatgcctt tagggacaag gagctgcggt 1080 gtcgcagtagagttgcaaag cggatagttc cgagaccttg agacacggag agatggccct 1140 ggaggtcaggggtgcttccg cggcgccttc caaagtctgt cgtccctgga gcaccaagct 1200 tccctagcgagtgttttgaa atgtttctgg gctgatgccc tcgactctta gggtttccat 1260 gctggagcagtagacacgcc tttcttttcc taagaccctc taggtctttc cgagagcgtt 1320 cctatgcgggctttgcgccc caaactagca tggaacagag ggcaaagcgc aactctgggg 1380 tgcgccctgaccgagaagcc ggaacactgt caagggtcct gctccagacg gcacccaagt 1440 ttgttcaacccccttaacct taccctacac ctagactccg gcagagagtg gggagtgagt 1500 gacctggattgccctgcggg ggcaggtgct ctagtgtgca gagagaagac cgtgacaccc 1560 ggcggctccgagtgtctgat tcttaagacg aagggcagcc acgaaggttt gtcctcgcga 1620 acctttcggcactgggtgag aggcaactcg gggcaactcg cgctgccctc gagcctcctt 1680 caatccgccccgcatctggt gacttgacac ccagctctag gagctgaggt gggaacacac 1740 tgtgaccccgccgcagacct gggtacattc tcgtcgcctg caaaagttta tagctggccg 1800 cctggcactttgctcctgtt tcgatcggtg cgtgccgtgg gttagagctt ccagcactgc 1860 gctccgtggcgacttagctg gggtgtccgg aatggtacgc gtgaccgcag aatggcgcga 1920 gctgccagagatccgtggcc agactggaaa agactggcgg tgaggaccca gccccgagca 1980 aacgggctttttgtttgggc ttccacttag ggacatgagc caggaactgt gtatcctctg 2040 gtcgaaagctgcagctccca cctcagggca ccgataagga gttgataaac tccagcgaat 2100 cgcacccgtcctggcccggc gcccgcgccc tgcgctggcg gatctctaga gggggaaggt 2160 attgagtccggttctagatt ccccattatc gcccaccaaa caactctgag gttctgtgta 2220 ctttgagggtagagaaggag aagcaagaga gagagagaga aaaaaaaaga ctaagattct 2280 agctcaaactgtaaaaccag ccagagggta atcacaaacc atcaaaaccg cgaagttgga 2340 cagcagagtagccaggagca cttattgaac actttattac tgccttgtct atgggtgact 2400 gtcgcgtttgtccgagatga gacctgacag taatcagtgt caagtgtcat cttgctctta 2460 ggtctatcaatccttgggag agtcagggat cagtggggaa agctatttcc atacccagct 2520 tgcagagactctttgccgag ggctggcagg gagtatagtc agagcctaga agaacgttgc 2580 ataatgacactacaaattat ccacaacagc ggccacttta agaaccagtg tgacacagac 2640 tgtgaacctcctaggaatct tcggcctgta gtgcagtgcc ccagtggtag ccaggcccgc 2700 agggtgtgtgtgtgtgtgtg tgtccaaaaa acagggcaag cagtgtaggt tcagtgtagg 2760 ttcagacttccaaagcctgt taggatagtg gttttgagat tctggttaaa ttgactgact 2820 tcttgggggcatcaatatgt agttgaaaat tgtgaagcag ggcatcaaaa gttaatgtcc 2880 cttaaatggctgcccagagc tttccagagc gcttcaggaa atcaatgtga gcagcaaggg 2940 ttcccggattggtgatacca cccactagtg gttcaagggc cggttcttct ccagggcata 3000 actgagcatttgtatggttc cttctggaaa cagcccttcc agaggccaga aggcttgttt 3060 tagtcccaaacatccttgtg tgggtggata tattccttca tgatagtcag gagccagtct 3120 tttatttgttggctggttag cagactgggg gagccacagc aagctctctt ggagttctca 3180 tcaacccagcaggattcctg attctaggaa gagccaagtg tggatgctct caagtgggga 3240 tttgtagaacaattaagata aatggaagtg aggtctttgg gtggccagga atccaagtcc 3300 aggcacaaagctagactctg ggtccaagta cctttcagga gtgcttctgg tatttggtgc 3360 cagggcaaagatgggcaagg aattggagat agctggacct tcaagttttc taactgtggc 3420 tggcttctgatggcagcaga tttaattgtt cctctgttaa aataaaccca aggtccatca 3480 gttttcccagccgaggagac aatgctggct gggcttcttg agtgaagctt atctcctaac 3540 tagtgttccttcaacatttg acaacagtct aacctggtag agcagtgctg tacgacacta 3600 tcagcctggcatttaaactg tactaaagat gactttctta gagttttggt ttttttgttt 3660 tgttttgttttttgggtttt tttttttttt tttggttttt tgttgttgtt ttggtttttt 3720 gttgttgttgtttttttcag ttttattgtg aggcttgaag attattttat aagcaagtat 3780 gaaagccataaacatatttt ctctttctgg ataaggctca aggaaggtaa aataatccct 3840 ctcggtacaaggaaaattag tgctgataag aatcagaaag ttggttcatt acaatcagtt 3900 agcatgtgaacagcagagcc tcccacccca tcccccccac ccccccaccc cccccacccc 3960 cgccatggcatgcctcttgc cactgaggat ttccaaagga ttagtgcagc tcaccatcac 4020 catacctccaccaccaccac caccagaccc ctgaaccttc aagacagagg tttactttcc 4080 tagataattgcaagagcctt gttctgagct gaggcaggag agatgtcaac cagtcagtgg 4140 cgctgctcttggtagttaaa aactttgggg aatcggaatc cattggttga gtacctttcc 4200 tttatttcagttaggggaaa acaaaaaaca aaaaccaaac aaacaacaac aacaacaaaa 4260 gaccccaacaaaccctacta ctcgaggtat aaaataaaac caatgcaagc accctcctgg 4320 ttcctggatgtagcaagttg ctttgtgaag tagaggacat tccacagcta ctagattgtt 4380 gcctggtgtgattaggcaat tttgcaggga tgaaagcctg aactgaaaac aggctacttt 4440 gtttacgttgtcaacaaagc ctctctgctg ctgtttctct gcctctgtgc atgtgtgtgt 4500 ttacatggccctgcccaggg ttctaggaat ctatgttgtt tagagagaga aggaaccaag 4560 tagttaattcctagtgctgt agggcttcag ccccctgggt ttgaaagtgc ctccttcaag 4620 gctgagccttttccatcttt gaggaaatct tttctgtgca gcatttactt gctttaaaaa 4680 ctgcaacaacaaagcaaaaa cggtcctgtc ctatctcaag ataatagggt cctccttcgt 4740 tttatcttgatgaatctttc cttagtccta aggaaagttt tgtggtacac aaactaccga 4800 gctttttaccaagttactga gtaagttctg ttttctccca gggccttatt attcagcttt 4860 gcacactccagaaacatgct ggtgagtctc acaatgacat ctataccaag atactaacca 4920 gtgggtctggaaaaacagtg ctgaaaacta agaaatgggg catctgttca ttactcccaa 4980 ttgcttggtctgatgggggt cagggtgggg ggacaggacc acccaagaaa ccgttggggt 5040 atcagccactcatgggtgtt ttctttggta gggttcaaat atttgaagct aaagaaaaga 5100 aattctgaatggtatttcag ttggtaaccg tacaccatta ttgaaatgca ttcgtcattg 5160 aggaagctggctctgtgtcc ctgagataat aaacggtaga tgctggacac cagcataaac 5220 tggatctcagagagagtgtg gtggtagacg ttagaacttg gagatgatga acataaaaat 5280 acggatgctccacaggcgtt ctgcaccatg aactttgaca catggacaaa atgactacca 5340 gttggttggttttttttttt ttttaaagat ttatttatta ttatacctaa gttcactgta 5400 gctgacttcagacacaccag aagagggtgt cagatctcat tatgggtggt tgtgagccac 5460 catgtggttgctgggatttg aactcaggac cttcggaaga gcagtcagtg ccctttacct 5520 gcccagccatctcaccagcc cggctaccga gtttaactgc ttgatgtaca cacttgtttt 5580 gggtggagaatttgcaggga ggacacttac tggcctccac agtgacaacc aaaggtttct 5640 gtttccactgaggacaacgg tggccttgac tgactccgtg aaggccgaga agggagggga 5700 gtacttttccgggagccttg taaatgagct tccggggtag taatagatct ctaaaattct 5760 atagagtgtatgttttgttc tctgggtttt ttttttttta atgttaatct caatctcccc 5820 tcttcatgaagagtgaaccc agagctttgc acgggctggg atagcattct ctaccactga 5880 gctggatcgccaccctttgt tcagctccta gtcgcctttc aaaggtggtg tgaagagaaa 5940 ataccttcttcctgcttgtc tgtctagcac ccaagagctc ccctccctga gttctaggct 6000 acaggctgcctgttcctaca tttcaatgcc agcgtcttgt cttgtcctag ggagtgtcta 6060 taagacatgttttccactat cccctgctat ctggagattc ttttgaacct tagaaatgga 6120 tgtgctcttgggagaaatga gctacatagg cagaaggaca agatggaaag ctgagcgctc 6180 aatcttcccgtgccagcgct gtcctcagaa ctttccggat taactttgtt aatccttgta 6240 acaaccctgtgagataagcc ctgttattat ctccagttca ctgatgggaa ctcaggtgca 6300 gcaaggttgctgcccgttgt cacaggcagt atatcttgga gctgagactc tccagagtcc 6360 acactctcacccaccgttcc ttgcctcctg gccttaccta aaaggcctat aaagcattta 6420 ataccatgtctgaatgtggt aagcactcta taaatgataa cttcttttgt tgctgtgttt 6480 ataattattataatgacttt tctcgtgact catcccttgt atcaaacctc acacacacag 6540 gcaggaactgaagacccaaa ggctattcct gaattcctgg aattacttat tttttttaat 6600 atatatatagatatttcctc agctaaagca ggggacagtt ctccactgcc cctggaagtt 6660 caaagctttcttcccaccct ttgctggtgg caagacttca agaaccgttc agtcttcaga 6720 ctccattgtgcggagggacc tgttaggagc tcttcctctg tggctccttt gatcttttga 6780 aattgcaagcataggaagga aaagatggat ggggggaggg gataaatagt agtttatgat 6840 tgtcgtgatcatatttttat agagtccttt ctctggtgag caccccaggc catataccaa 6900 gagccttgagatcttcttga gtttcctcgc ctcaaaccat gattgctttg gaacagttgt 6960 gggttgtggtatgggtttgg ggtggggcgg ggtgggatgg ggtggggtag ggtggggtgg 7020 ggtggggtggggtggggcat gtatgaatat atgcagctag caacacacag tttttcaagt 7080 agagtctatgtgcaattcac tgacattttt ctggtcctgt tactcaaaga cacagaacca 7140 atctgaacgtccgagactga gagaaatgtg gaggttattt gagatcccca gaatcagaga 7200 taagcgtggccgagaccaga gtgttatgtg cgtctgagtg gcctttcttt ccctaatagc 7260 tcaagaaatactggtttgct tatggccttg ggctgtttcc agaggccacc agtacatcct 7320 ccaggctcctgggataaaga gtgttgcctg ccgagaagca caggattctt taaagttagg 7380 agtggtctgtatgccagacc ctaggaagca gatactttct catacagaat gagatttcat 7440 gatgaatggcttgttggtaa gtaaaacaaa cacatgaaaa cgtgtgtgtg tgtgtgtgtg 7500 tgtgtgtgtgtgtgtgtgtg tgatggctct gcacgtaaga gcatttgctg atcttataga 7560 agccctgagtttgactccag cacccatgtg ctcataacta cccctcagta tagttcctag 7620 ggatctgacacccagttccc acagccacgc agtcacatgt cacacactta aaaataacac 7680 acacacacacacacacacac acacacatgt atgtatgtat atccgtgtat acatctgaca 7740 tgttttaattccacaacatc atggaaatag acctttagaa ttttcctata tcccccccag 7800 tggctcactacattcaccat tctagtggca acttccgatg tcataggtaa gaatcagatg 7860 attcatgtttttcatctggt tagactatgt acccaaacat gtatacacat acacacacac 7920 acacacacacacacacacac acacacggat ctactgtgct gccctattga ccatgatagt 7980 ctagagactgacccatgctg ttatattggc tacccccccc ccctaaatgg caggctacag 8040 ccactggcaattttagatgt taatgagtag tgttagtggc ctcttttggt ctgcccttgg 8100 tttgaagtgcacagttagtt ggcctcttaa ctggaaagtc agtagtttgc taagaataag 8160 aggatacataggctccatct aagaaagtta cgaatgcttt ataaatcatt taaaatacac 8220 cagctacagtatatggtttg gaactggcaa taatttaaag cactatgttg tgtgtctagc 8280 catgaagggttctgtgtctt tggatggcat atcactgtta acaaacacac atgttactca 8340 taattgaaagcgctcagcct cttgtcacca tgtaaggttt gtcctgacct gcctgcctcc 8400 tggaggggcgggaactggag gggctaaaga accaaacctc ttcatctgaa gccaaaccta 8460 ggcgtggagaggctgggccc agaacaccat cttgggctcc tggtcttgtg ttgtgttgta 8520 ggaaggggccatagccagga acctaaagga gcaacttagc tgcagcacta accccgctca 8580 ttactaatgagtaattacag caaatattta caggtctcct tcctaccttt atgacttcct 8640 gttgctgaagcactttggag tgttgagaaa gattgttctg gaacatacgt catacatggt 8700 tgtaggaaaagagtgcctct aagcgcacac acacacacac acacacacac acacacgctt 8760 ggttttgtttatgaattcca tgtgggaata acaaaattac accttctaga agctgagaat 8820 tttacttttcaaaacaaggt ttaaaaatca attctcaact gctggtttta aagcaccaat 8880 aaggctattcatgttcataa agtaaaactt gaggttaaat cataggcatt gcctaaatac 8940 tttttaaagcactacaatgt tttatgaaac tgtgacccct cacagtgcct tctgagtgaa 9000 aacagaaaaacctagtagaa taaatcaagg tctataaaac aagccatgtg ggcaattctc 9060 tgagttccagaagccctggt aatgtattta gggtttgttg ttgttgttgt tgtttgtttg 9120 tttgtttgtttgttttttaa aatcggtcca gtgagtggct ttctcagtct gctcctgtga 9180 gccactgtgccctgtgtttc tctctggtgt gtccagcagt tcccaggtgc tattttagaa 9240 tggcagccttcgctccagcg ctggctgagg ttgatggagc taattttggt gtctagggta 9300 aagggagcgagaaagcctgt gcaggagtca aaaaggcttt cttgtcccgg gctgagagca 9360 gcgtggtgttaccagacaca ctacatttca cctttagaac ctttgtttgt gtacttgcac 9420 aacaaatgtttacctagata ttaagataaa ggaaatatga aggtcccatt tcactggtga 9480 caagggagcgtctataattg taactgtatc accgtgtagc aagcaggagt cccttttact 9540 ctatagccaggtcttaaggg aattaatggt gggtgtggtg tacaagccaa ggcccttctc 9600 agtttctgtttctggcttta ttgctgctac tatgttttct tggctcagcc aatgttttgc 9660 cccacccatttggtgaaggc ctttgtggtg gtcaaggatt cagagggaat tctaaagcac 9720 tctgtggccccactctggag tcatccgctg ttctatggaa accagttagc agaccctggc 9780 accatcactcttttcctagg ctctcagaaa acgtttacat ggtaccaata cgaccttgtt 9840 tcaggccttcacgttgtctt ggaagcacag caaattttcc ttgtggcaag agggttccat 9900 gaggacttgggggtttcttt gaagatcccc caaggattag ctaaatactc agtctgaaga 9960 tctaagaacctcactcaagg gccttcccat agggaagcta cgaagcaggt gactgctgga 10020 aatgagggtccccacactcc agctctctca tctgccccac tcagtcactt acggcacctc 10080 ctgagctccccacaagccta ttcctctgcc acacaggtat tgtcacagta ctcgttcgtc 10140 tctggttcttagtccatgct tgaatgcttt ttccctcttt ctgcccaaat atctcacact 10200 ttgtcttatcatgaatgagc ctaagctctc tgcctttaac agaaacccat agcattccca 10260 ccctgcctttccctccattt gctgttttat acttctttgt tgtctaaaga cattttattt 10320 gttcttgcctttccacgttc gttcttgcac atggcacaag agcagggatg tctgattttc 10380 tgatgtggccaataaatgct gatccttttc atttttctct gaaatcatgg ccattagaaa 10440 aataaatacagaatgacttt ggttttttgt tttgttttgt tttttcgaga cagggtttct 10500 ctgtatagcccctggctgtc ctggaactca ctctgtagac caggctggcc tcgaactcag 10560 aaatctgcctgcctctgcct cccgagtgct gggattaaag gcgtgtgcca ccactgcccg 10620 gcttcgaatgactttgttga tgtcccactt tctcgggctc agctgtctcc atccacacga 10680 cttcccttcacctgtttctg tgaacaagaa aataattggg gttgggggag gggcaaggca 10740 cgggccaggtgagcattatt tgccatcaga tgcaaggatc aggcaggtag tatgccaatg 10800 ctgttgtgcatcaagaatgc atggtagtct aggtaaactc cccagcttaa ctccggagtt 10860 aacagctttccaggtgggaa attatgcaaa tgcatccgta tcagctgtcc caaagggtct 10920 ccctggaaagcagacctcct tcagtcagca gggcattgtg tagccctgta tccatctgat 10980 cttctgtctgaatttttaaa tttgcataat aagtttaaag cttgctttct atttccattt 11040 taggtgttctgtagttgtct gtaggtggaa ttcaggaata tggggacctt agttcaaagg 11100 gatggaaaccacaggcagaa acactgcctc agatcagctt acccatcctt atttgataat 11160 caccacgcaggtcagaagag gtttgtggta ctgctggcct cgagtataca tgcttacagg 11220 ttatttgctgatttggggac tgtgctgcac agagagatca caccaaggca tttgatgact 11280 ctggtctcttcaaatgactc gtttaaacca gtatgtggca gtgtgtggca ccaaggcacc 11340 tggctgcatctttaggatgt tgccatcttg gtgtggagta aaaactaggc cagatacagc 11400 acagagccaggagcagaaaa tggataggaa tcgagagctc tgagttgtat aaagttaaaa 11460 aggactgggctggagaagac cagagctttg agagaagagc attaaataag acttcggtct 11520 tgaagattagaatttgggtt tgcacaggca gcaactgcag gctacgctgg ggatgggcga 11580 ggcagtgcagaccgcgagac ggaaagaagc agagtgcgca ggacagggct tggggggtgc 11640 tcctttctttcaactcggaa tcagtagatc ttatattagc agagatggag cctgggcctg 11700 gactggagaatctagggaca ggccctcttc gagcatgttt gatggaattc tcagcctgcc 11760 ttttttggagacattagaag ctatttagag caattttatt gtccttctcc cacaaactta 11820 gctagagtgtgcacacctag gagtgattgg cagaagccat gatggcctcc aaaacaatga 11880 cctcaaagcgctgaacccat agatcttagg aaacccactg tcattccgtc aggtatgtat 11940 ttctttgtggcttgcatagc tgcttgggag tgataaatac tagagttcta tcagaatggg 12000 aacctgaagggggacggctt gctgttaccc taaaattgcc ttgcccttca ttcccatgtt 12060 ctcactaggaggccaaagcc tttgtcccct tggggacttg gtaaggtcat ttgtcatatc 12120 ctttcttgccatttctcctt agagaaggat aaggcttctt tcggcaccta gcacgtggag 12180 ggagttacacagccaaggct gagtcaaatc tttagtctct actggtagct cttcatccaa 12240 acccagcccagagcactccg gccaagccgt tctgtgacct tctctttaag acccagatct 12300 ggttgatgttctgagtctga ccatgccaga agagtggggg aaaaaaagaa tcctctttcg 12360 gagtcacctgggtaggacag tcagagccct ttcctctcca caagcaggtg tgaccttttc 12420 cctggaggaatttagaagca ctaaggtcac agttggtcaa agtgggccag gagttggtca 12480 aagatcccaaatccctggac aagagcccac atcagggaca gcagaggcca gcaactgatc 12540 ctagttacatgagtttaccc tgcccagtgg tacttcaagg gagggaacgg ctcctagatg 12600 gttttgtgttaaactttaac ctcacaatga caactgtcat gtctcaactg ttagttctgt 12660 cccctgtggttgtaggacgg aggaaccact gctcagttct ggcaggctgg ttaggccagt 12720 ggtttgctcaggttaaagcc tgagcccaga gaagagtctt ctggagccaa ggagccgtaa 12780 tcgcctgccagaaccacata gggacagggg acagtggagc tttgcagcac agtacagact 12840 ggcccttatccaggagctaa ctgagacctc gggccatccc ttctggaagc ctcagggaag 12900 ctctaagaaaagccagaaac caagaccaga cctgaccaca catactcaga tctctccaca 12960 ttataaatgcgagtgtagca tctacattcc gatagcttct tacaggggtc aggaagggaa 13020 aaggaagatgtcttagccaa gtttgcctgt gactaaacac cagatagcta gctccatgtc 13080 tatgtatctgtgttttcttt cctaagtgtg tttcttaagg tttaaaaaga tgcatgtgta 13140 tgcccgggtgtttgtatgtg taacacatgt ctcaggtacc tgcagagacc agaagagggc 13200 gctggatctcctgaagctgg aattacaagt ggttgtgagc catggatcat gggtacttgg 13260 aactgggcctgggccctctg tgctcttagc tgctaagcta tctaccccac cagcgtctgt 13320 gttcatgttcttttccaaag taaaaggtcc ttaaaaaaac aacaaacgag caaacaaaca 13380 aacaaacaagcaagcctttg cttctcctga actcatagca ggttcttcct ggccttggtc 13440 agtgaggggctaggcccggg catagctcaa gccagtgtgg ttctcatgtt ctctagctca 13500 ttccaggctatggggagatc cagaggacta gcgcgctcct agtgagtgca ctctccatcc 13560 tgagccatctctcaagcaca ttagattctt ttctctaaat cagtgggtgc attttagttc 13620 tggccacctgtagttgcttg tgtggggaca agggtggtga catcacctta attttcctgg 13680 ccttgggtccccggagccct tgtttatttc ggggagtgac aagctttcac ccacttgaat 13740 tccttcgcctccaaatagcg tcagaatgac cacaagcctc ctgtgtttct tcgctttctc 13800 gggttttgccagattctaaa tgccgtcagg gccactggct cttgttttat gtccctggca 13860 aagctggccttcccatgaag ttcaaagccg ctttcaggca tcttgggagt ctgggagact 13920 gctttcgcttgctctctgct ttgtgcggac ctaggttgga gatgtcacct ctgtctgctg 13980 ctctctgaatacgaaccagg gaatgttcta gtaataccac gtgctttaaa tgtatatttt 14040 aaaagcacactttgtgagta ttatcttaat ggaaagaact ttgaaaagta taaagtgcaa 14100 accttctaggcattgtcatt aaggagcaga gcaatatact cattaggtgt gttattacct 14160 cttaaaagtgaaccgcctgc agacaggagg aagccttgag agaggctaac aggagactct 14220 ggtctccaggctcctaccgg tgggtccctc cgcctgggct tttgggtcct gtggattctt 14280 gccaatcgtggcatttagtg gataccccaa gactgaggaa atctgaagaa agtcctgaca 14340 agcaccagatcccaacaccc ttctgcttgc tttgtttccc ctcacctgtg aagcaggaga 14400 gggcacagcccagcactcac gtcaaggacg acacatcctt ccgtgtcgca caggaaccag 14460 ggctgcccaggccatagctg ctcgcctctt cctcgttcca ctattttatg ccaaagagag 14520 gcattgacaacctagaaaca ggtgctactc taaagagacc ttggtctcct tgaatgcaga 14580 ggcctggctgtgcttatacc ctaaccagga gacttgaccg gtcactcagc tctggcctca 14640 gacctatcatcaactgtaac acatctggac ttcttacctc tgagctctct cttccctcag 14700 ccccacccgggagactggcc agctgacttc aagcggtcct tctcagctta aatatcacct 14760 cctcagggaagcctttctcg acgtccttcc attgacctct aatcctgttc catttgtctt 14820 ctaaacattttccacgtgta atttacatgt ttacttatgt ctaccatgcc cctcccctgc 14880 cagggtgaaactgaaggtat ggactccaca cgtgccttgt ttaccactga gggctcagcc 14940 tttagaatggagcctgcatg cactgctgct ttttacagat gcgttcagcg aactcgttgc 15000 tccgatattgctgggctcta cattaccacg gtttacaatt gtccagtgtt ttcctaaagc 15060 tgattttgtttttgttttct gaggtagggt cccaggggct aggatttaca tgaatgcccc 15120 catacttgttctttccaaag ctcttggtgc taacaccaag gaattgtcac tttttagcat 15180 atggatgaggcagttgagac actagagtat aatgaccatg ccagggtctg gcagtaccta 15240 cctgaacacgttcccagccc cagactattt gcaaagatcc acgctgcctc tcttggcccc 15300 atagttttctgttgtggcga tgttattgtt tgtcatattt ggcaatgttt accccagaga 15360 agtagggccattgtgtgctg gtagcgtctg ggaatgcaca gccaagtccc aggggaatgg 15420 ccactgctgttcttaccaca attagaaatt gtcaagccag gagcagaagc agggtgggtg 15480 ctgccatacactgctggttc tgcctctcca tggggctggg gtgagggtcc tagctccgca 15540 gccccgtgtgtctccttgtc ctggctctcc cactcacatc gaagtgtgga ccttctcctg 15600 cagggcgatgtgctatgctt agtgaatttc ctgagaagag gtaggcatta gctggctaga 15660 tgaccacctctggcctcaat tattcaatta ttctaccctc tccaaaatga accagtagat 15720 gggaaccagaccaggtaacc ccaaaactct ccaggttcta gctccgctct gaagggaatt 15780 tccaggggtctggcctcctg tttgcagatg ctgactctgg aaagagcagg ggaagttgga 15840 ggttgttggcaggggctggc ggacctcttt ccatctcttt gtaactcttc ctctccaaaa 15900 agatattccatcccatcagg agttgctgtt gggcctggtt cagtgcagca ggatgaacaa 15960 ccgcccccaaaagtcagctg aggtctgata tgtgatatgg tagaaagctc ccaaaaggag 16020 gccatgcctccatctccctg atgcaggctt ctggggtgtt ctgatgccat taacggacag 16080 gggtcagacaccaagtgcct ctgtctaggc cttcgttttc atgtctggca ggtgactgtt 16140 ctgtgcccttcaattgaaca aggctgcaca gatgtaaact gccacagagg aagggcacat 16200 tgctgctatgattcctgtgt acgaatgttt ctggcgtgct cacacctggt agtgacatga 16260 actgattgacacttgcagcc tgcaaatacg gtcctgcaac ctgaggcacc aagggagaag 16320 tcagctaggaagcccgtgag gccttaagtt gttgaatgaa gtcatgctgc acagggtggg 16380 gggtgggggtgaccgtgctg caggatagag gtgagtcaca gtgcaagact gttggggagt 16440 caccttgaatctgagccaaa aaagcagaaa tattgggact cgtttatcag ccttctatca 16500 ggtacatcaagttctggatg gccacccact ggccagcgac atgatgtgga cggctctgct 16560 ctaccgccctgggaaggctc tctgctggct cttgccccgc tgagcaaagt ccgcttgttc 16620 gctggagttcacacagactc cttgccaggc ctgcccagaa tcctgtctcc tctgacttcc 16680 tgtgttcttgcataatattt ccttgcctct tgaatggctg gccccagtgc gggggcagct 16740 cactggctctgctggattga gagtaggatg tggagggagg atgggtgata tttggatcta 16800 atccgtgggtgctccgcgtc ctggtggcag agcccctcaa actttttgat ggaagctttc 16860 agccagaagggagtgagaga gcattgcaga ctgtatacac acactcgctc gtgcacatgc 16920 tccctggcttcttatattca tgaatcatct ccctgggaaa attgttgaag tagttcaatt 16980 gttttcttctggtcataaaa atatgaatta ttcccatata gtcactgtat aagaagccta 17040 aaagtaaaataaatatatat atatatataa ataaaaaaca aaaaaatgta ttattgaaag 17100 ataagttccattaacagtga atatagtttc tcctacgtca gaaaggctga tccccatggc 17160 tatgggataccagcaatatt agctgtatta agttctgctt actgatgcat cttcatgaga 17220 ctcccattcactacccgtcc aggctcaaaa agcaagtggt aactggccca acaattattt 17280 ccaaacctgaaaagcaatgt ctcccctgaa gttctgacca gtatgctctg gagtcccaga 17340 aataactgcaaaatcaaagg ctggtgcctg tgtgagcctt tctgtaggct tagagtaatt 17400 cccatgattacacaggagaa ggctgcttcg atgacagctc tgagggctgg gcctggtctc 17460 actttggggaaaaaaaactc catctatcca cagggccgtg tgtgagttaa gccaggagcc 17520 cactggctggcagagcaagt ttaactttgg ctttccagtg ctcaagattt cagggaggct 17580 ctatttaaattttacttggc tgtcaccctt ctgaaaattt gtagcactat ccgtatcttt 17640 aaggaagtgacctttccccg tccttgtcct tgttagcatg acatgagaac ttggaagcgt 17700 ccatggtgacctgcttccag tttgattatt gaaaacaaaa caaaaccatg catagatccg 17760 gggtttctgacttacctctg gaaactgtac tttctacagg gtggccatga gagtttgcag 17820 gccacctgctaaaagttgac aacctgagag tctgcagtag acaacacaca cagcatgctt 17880 ctgtgttggatctgagtgtc tcctgcatct gtctgttttc tttgcttctc ctttaaactg 17940 ggttaaccatcctccatatt ggtactgggg atagatagca cccagggcct cacagattgc 18000 tagaaaagtgctctaccaat gagcaacgtc ctctgctcaa tgagtgtgtg tgtgtgtgtg 18060 tgtgtgtgtgtgtgtgtgtg tgtgtgtgta aatatcagaa acttggttaa ataaatatat 18120 gtaacatacaatatacttat ataatatatg tgtatatatt atatatctcc atcccaaagt 18180 tctctgtttgagactggact taatacttcc cgttgggaat tgcttataag gttttacttt 18240 ttagtaaagaaaagttgttt gtttcctaaa ctctctggct taaacttttt acttaagtca 18300 aaaagcaagagaatacctct cggcttgaat acctttattt ttaaagctca agagtctttt 18360 ttaaaacaaaaacttgctac agaacttggc accaggggac tgcaaggatt taccttattt 18420 tgatgatagctgcctgcaaa cgatctatta gaataatctg cataattgca gtctcccctt 18480 cagtttattcagcctgcact ccctagtcta gatttactgg ccagactgtt attacatcaa 18540 attccttgatgctgtattcc tgaggactga tggaactaag agttacagag aggaatcagc 18600 gacctttgtctgcttttcag aattcttcag aacatactga tttagccagg ggcttttgcg 18660 attgctctccttactgggtc agtacctgat tttgctgggt ttctggccac acatgtgcgt 18720 ttgggaagcaggatagatag ggcgagaaac ctgaattggc agtcagagtt atgaaacagg 18780 actttgttacaattgatagg tctgtacagt ggacgcacag cagctgttgg catggtaact 18840 cctacgtggcagagtgcatg gagcccgcag atgactttag cagcgctccg tgttaatttg 18900 ataaatggcttttttaatag tagtttgtgg gctaatggaa agattgaagc gaaccctcgt 18960 taatggaagggaggatggag attttgaagc cactgggtga agtcggagag tcccagccaa 19020 gtctccattttcagctcagt gtctcttttc atgtgcctga agtgtggcaa accaaagtac 19080 agtagggagcctgccttgag agtaggcatc ggccctgggc tccggcttac gagtgaagag 19140 gcttcagggtccttattcaa tacagttgct ttgtgcaggg gcttagctta gggccactgt 19200 aagaagtctacattgtgttt tctttttcct tagtggtaat ggggcagggg ttatggtggc 19260 cgacaggtctggcatattta gccgtttggt tggttgtatt gtggttattt gggttctcag 19320 aggggttgtttatttttgtc tctggtttgt ttgttgacat agggtctcgt aacccaggct 19380 ggcctcagatgccctacata tatatatctc tgaggatgac cctggacctc tgatccttct 19440 actgccagctccagggtgct ggggtcatag gcgtgaacca ccttgcctag ctaatgtggt 19500 atcggggattgaacacagag cttctgcctg ctagatgaca ctctatcaat taacggcatc 19560 tcctcctggtttcggccatc tttcattgtg tgagaataca taatcacacc acactgctgg 19620 cccactcaaggagcctctct gcaacccaca ctgaagctct gcgttcctta tatagcgctc 19680 gacaaacacctctgggggat ggatctcctc actgtgttgc caaagcaaaa cctggggttt 19740 tttgcctttctggcaagttc ccaggagacg cggacattac cggttctcag tccacatgct 19800 gagaacttggtgctttacac taatgcggag aaaattggga acaagacata aagaggccaa 19860 atgacttgcaccttaatggc taaggatgat ggagcttgaa cttgaacgtg gctagggcct 19920 ctaaggcacctgctgtctct gtcttatcag caatggcaga agtgcagtgg ctggagcatc 19980 ccaagggtaccttctgccat cccaagggta cacggtgtat taatctgtca cagtcaagtt 20040 cagaggtggcctcggaagcc tgctgctcac agccttctct gcgtgcgcat tggagttggt 20100 tttcccctgggctaacctac aatggagaca gtaccaacaa acccaccaat gcagctgaaa 20160 caaaacaagactttatccgg gcattagtct aggccgccta ggagagaagt ggtttgtggc 20220 ttagatttgtaagggacgtg gcatctccag ttgttctagg ccagaacaga tcatcacagc 20280 tggagtctttgttcagatct caggcaaccc agggtacttg agaagtttaa gacagatagc 20340 attactagagaggtttgttt ggttggcttg ctttttacat tttcatactt tttctcctta 20400 actcatactccaaacattcc attacatggc gctaggtgta gtgacggata ttaagatggt 20460 ggccagtgaatacttgctaa gaaaagtagg ccaaaggcat ctgtgtccaa atatgctgaa 20520 ccgctcagcccaagagcggg agggatgatc aagacagaag gacaggtaac acctgggatt 20580 cacacaacacattggctctt aaagtcacat tttcaatgtc cttaaaaaac aaaatgcaca 20640 gacacagacacaattaaaat aagtcttaaa agaaaaaatt taaaaatgca aaaattatag 20700 tggctactgctactgttctg tccccaggat gtcttccaca aagagggaat ggaagccaaa 20760 gcagggttttgtgtgcgtcc ggagcctcct gaccaatagc tgtgattctt ctggctttag 20820 aaataacccaatgccatctc tccagtggct ttgccaaccc acatgatacc tatttctcat 20880 tacccccaataacgaatatt tagtctgtga ctgttgtgta tacagggtga tctcgtctca 20940 ataactatttataagcaact taaaagcaag ggtgggatta ggaggtatat atagctcgat 21000 ggtaaagtgttctttagcat acccaaggcc ctgagtttga tccttagcat aggatcagag 21060 tgaaaggggcaagggcaagc aagatgtttg tggtggtgga ggtggtggtg gtggtagtgt 21120 gtgttactgtgtgttactgt gtgtgtgtgt ttctctctct ccatcatgtg tataatatgg 21180 catgtgtatagtaaatatta catgtaaata ttcaccatat taaaagctat ctgtaaatgc 21240 aaatttaatttgtgaagaga taaagcttcc aggtccattg aagaggagag gtggctctaa 21300 ataggctgttgcagacaaaa cagacaccgt caggagcaca gatgctctac tttggctgtg 21360 acacaccctgtggcagagaa gactggggtg agaatgaaaa atggacatcc ttgggcaact 21420 cccagatgccatgggttttc cacatcacat ttgatcttat aggtaaaatt gtctttaatg 21480 ctggggatccagtgcaaggc ctcacatata gtaagcaagg cttcttccac tgaccacatc 21540 catgacactcgctttttttg tttgtttgtt ttgttttgtt tttcaagaca gggtttctct 21600 gtgtagccctggctgtcctg gaacacactt tgtagaccag cctggcctcg aactcagaaa 21660 tccgcctgcctctgcctcct gagtgctggg attaaaggtg tgcgccacca caactggcga 21720 cactagcttttaaagtcata ttctaaaaac tactacataa gtgggtatct gcgcacagct 21780 tgtaacagacttcacaagag gggcagagtc cgtgattcgg agttgttttg ttaaatgtca 21840 agaatacagaaacatagaac tggttaatat ttctgctttc gatgcaaatg ggttaggtta 21900 gagccaggctcatggccttc cccggtctta ctctgcctca tgtagcttgg ctacaggcct 21960 tgggcatagtgttggattaa gcagagaggg tccctaatag gtatgttcag gaaataggta 22020 tgttaaataaaggaaatggg gtttggggtg ggctgaactt tcctgaagga gcaggaagat 22080 tttctctagtcagatctttg taagagcctc cttcttactt acaattaacc ccccccccac 22140 acacacacaccatggaagat gtcctgacac cctatggact cagggttcac tgcttagggc 22200 tctctgttctatatgcttaa gtcaggatct tagagataag ggttgtggaa accaattcct 22260 gagttacatcacaatactaa tatcctttgg aagtttttag gtcattacct taggaaggga 22320 agctattttgctattcattg gagatgggag gaggaacata aagcaaaaat ttctgctgat 22380 gtgggtgtttgcttggggcc aaggtactgg gaggggcacc aagggtgtgc ttcctttatg 22440 tttgtaaaagccctactagc ctctgtttaa gacggtcctg taggtaggtg caggagggca 22500 tgaatcattttgtgctgcct tcctcttggt tcagcagagc ccagcaagtc agatgggtag 22560 aggtgacctatctggcctgg tcaggctttc cattggtcag cagcaaaact gtgctctggg 22620 cactgatgatgccaggcccg tgctggggcc catgctggag gatgaggtcc acaagccagt 22680 tcctctgtacctccaaggct tacaaaaccc cagccactgg gctgtgcaat ctcacttcaa 22740 atgagtgttatgtccaccca tggccgtcac acatgagcat ttcagtggaa agagactgaa 22800 attctattgccatggacttt cagaactcat gctcgatgga gatagaaccc accagtgtat 22860 taggttcttgaaagctacct cctgcgcatc atttaaatcc taaaaagata atttccaatg 22920 aagagaaactgattattttt tgctagggac aggttggctt aaggtgggtg ctattcgaga 22980 tgtctgagacctgagggatg accaggatga gaaggaaatg gtcctacact gggctaggtc 23040 ctccaggcaacccccctaca gagagcagtc tcctgcccgg ggtggatttg ggaccttctg 23100 aaatctttgtggtccaccag tagggaatca acttcttact acggagagca gctggagacg 23160 taagcataacgcctttccat tgtcccggcg tgtattctca agtgggtccc ggcttcctgg 23220 aacggcttcccttgagtgtg agggctgaga tgagtctgcc gggtgatgaa tgggttcagg 23280 aaggagtggctgcatcacct gcctggggga tgagcaccca cgtgacttca tggttgtgca 23340 agaattgggcaacgtttggc cagggtggag aggtcttggc aaaggcagtt tcactcctaa 23400 cagattcctatctcctccat gggggaaaaa caactatcag gagatccatc tgtacagcat 23460 tggaggacgttgatcgcttc ttcagctgtc tgtggccttt tatttgctaa gaactcatgg 23520 attgaagacctcagaagatt aaggaagata ggcatccctt tccttcctgt ggcagctctg 23580 gagaaggggagggtgggtaa aaggaagaca aggtgggagg ccatcaatgg caggacgggg 23640 agaaatggttttagagcgtc tgcagagtag tcgcagagca gagtggtagg cttgcaggtt 23700 caagttgtggatctacctct tagctgaggt atcttgataa agtactttaa accctctgta 23760 cctatgtgtttaaagtacaa gatggaaatc attgtagtat tgaatcagag ggtagggctt 23820 agtcatcatgaaggttggtc ctagttccaa gacactagac acaagtggaa gggctgaact 23880 aaacttgggtttttgactta ccactcttca ggtctcaatt tcttcatctg tacagtaaag 23940 agactagagcagattaatgc taaggttttg tgtattctaa atgatatgat tccatggttg 24000 aataactattaagtgtctgc tgtatgttcc agcactgtac ggggcatgcg tgaataggga 24060 tctctttgtccttaagatct tgtcttactg gggaatgttc actaatacac aggagaacat 24120 ttaattcacaaatcaactca caaattaaaa aacattagaa gccagacatg gtgacgcacg 24180 cctttaatcccagcacttgg gaggcagagg caggtggatt tctgagtttg aggccagcgt 24240 ggtctacagagtgagttcca ggacagctag ggctacacag agaaaccctg tctggaaaaa 24300 caaaaacaaaaacaaagcaa aacaaaaaaa aaacaaaaca aaaaaattag aaattgaaaa 24360 cttggagcattttggctgga tagatggttc atctgtcaaa ggcacttggc tgctctttcg 24420 gaggacaggcatttaattcc cagcacccac atggctggtc acagttgcct gtaactccaa 24480 tttcaaggaatctggtgccc ttttatggcc tccatgagca ccagttaggc atgtgatgca 24540 tatatatgcagacaaaagat ctatacagaa gtcaaaagta aataaattaa aaacccaaat 24600 gccgagtaatggatctgaag aacattggaa taagaaattt cactgtggac cagagtaggg 24660 agacttgacagcagctatgc accttgtcac atcccaggaa cactagcatt aatactgaag 24720 ctggagaaaacagcctccta tttgaggctt agaccaaatt ttataagaag agtatagact 24780 gaaagtatgatgtggtccaa actggtttct catatattct tggatgttca tcctatcaga 24840 acaacgtatctggcacacgt gagaattcac tttccaaagg ctgggaagat ctagggctct 24900 gcattgttagcttcagcagc acgtagtgtt ctcagccctg cctctagagg gtagcgcaca 24960 ggcaatacccatcacacaat acccatctta cctatagctt ggagaagagc tttgagtggc 25020 cttatgcttcctgagccaga ttcttctaag ataaattctt ccagtgccta aactttgacg 25080 acattgtgggaggggaatat cgatacacac ccctgaagtc tctgcatcta catttggagg 25140 aaacttagcaaccccttcaa aggtgtttca taataaccaa accatagttc ctcatgcaaa 25200 ttggctgatccaggcaacaa gggaatattc ctaatggcca aagtagtgga gttcaggtca 25260 tctcagtgatagagctggga agaggcttac gcaggtcttg aaaagtaaga agaggccatt 25320 ttgggagataagatgggtaa ggcctcagct acctaccagt ttccaggcca cctctcccaa 25380 tgcacacatgcgcgcgcaca cacacacaca cacacacaca cacacacaca cgcactcttt 25440 ctcacacactcttttacaca tacattcttt cacacacaca ctcttacaca cacattcttt 25500 cacatacacacacactcttt ctcacactct cttttacaca cacattcttt cacacacaca 25560 ctctttctcacacactcttt tacacacatt ctttcacaca cacactctct ttctcaaaca 25620 cactcttttacacacatatt ctttcacaca cacactcttt tacacacaca ttctttcaca 25680 cacacactttctcacacaca ctcttttgca cacacattct ttcacacaca cactctcttt 25740 tacacacacattctttcaca cacacacaca gacacacaca cagacacaca gacacacaca 25800 cacacacacacacacacaca caccacattg caggtagtca gtgcatttgg atgtggttct 25860 ttatttccagacaggaagtg agatgtaaat gacagatgag gtgcatgaac tctctggcct 25920 cacccagacactgataattt cccatcatct cttgagcagt cagtgatggc ctggctcgat 25980 agggcggttcatgacaccct agcttcagat cagcagattg cagcttgttg ctgagactcc 26040 ttcctgttacagaccacaga aatcctggtg acatgcggcc cattttacct tgtgtaaagg 26100 cacaaggacatgtcacgctt gccatgagaa cacccgttca cacaggcacc aaagcagtag 26160 gcaggccagatggagtcaca gggttcagag aaggactgtg acataatgct gaagccccgt 26220 gttggggacagatgtctctc tgccttccag gaggcggcag taagcgcttc ttttccaaac 26280 cctcctctcatcccggtccc ctcccctttt cgttcataaa aaagttattt tcttccaaat 26340 aagcaattccaaaatatatg aaataaacgt tagttctaat gagcctctgg gaaagtgctc 26400 acctttgaactcggccaagg attatgggga aaagaaaaag tcgtaggaac ttgatagagc 26460 gttagagcttcctgggtttt taagctgggt tatgtattgc atttctttgc cttaataagg 26520 acggttccagaactctgccc tggataattg ggccatgtct gatagtagag acccaggatt 26580 ggttactggattagggatta ttatctgggt gctaggcaac aattgggtag gaggccctgt 26640 ttctagaatgttctttcttt ccaaggactc agaacctttt tttttttttg atggtccctg 26700 tggggagtctgagacctagg aagaaacaag aggatgttta taggaggccg actgctaaag 26760 gggagtaacactcaggaact gtcctgctga gacaagctta ccccccccac ttccccgaga 26820 cattgctgcttcaaataaca gaaatcattt tctgaaagac aggctttcag tctgggtcgc 26880 ctctggctgcttgtatggac tcttcacatc tgaatttccc caccctcctc ccccagataa 26940 gaagtttacttccagccatt ggcacaaatc atccctaggg tgacttgaac ctgactaagg 27000 acagctctcgaaaatcctga taaggtctcc aaacttctat gccctgtagc agtaactaac 27060 cattcccctttctttaaact cgtccatctt gctttcattg tagtgttttt cctcatgcct 27120 aagtcaaatgagctctgtgt ctcatcccta ccactctcga gggctgagca tcaccagtgg 27180 gctcctcctagggaccagag atctagaaac acagaggacc ttagctgagc tggaggtacc 27240 acttcacggcatgagctgtt tttcctcaat tttccttcct aggcctgcac agatttcttt 27300 actggatggataggccccca tgcataccca gcctacctcc agcccagcac ctgctcagta 27360 cacttagcctgtaaatacag tcatcccaca aaggacattt tctctggtgg ctttaaggtt 27420 tgacggagagttctctagac ttggcagctt agctgtgacc tcaggaatct cggtgctggc 27480 aaggctaggtctttgcacta acgtggctca gtgcccatgg agataatctc ctcttgtaat 27540 gggtgcacaatcattttatt gttgaagcaa taggaacgca aaaacagaag gaatcccaca 27600 acagagcttgcccgtggctc agtgagcctc tctgctcctg gctgagggca ccttgggaca 27660 cctcaagttcaatcccaacc gccaccctga aggtcagctg attcataagg tgtgggctta 27720 tggaggagcctacacccaca gcctgagcct ccccaggcct ggcactgccc tgtgttctgg 27780 ctaacacctcctgtttattg tttgaaactc aaaagacaaa accctccagc aacctcctcc 27840 ctttctactctgagataggt accccctttt gtccctatta ccactgactt ctgcagtagc 27900 agcatttggctccaacactt gagtgctcgg taggcatcag acacagttct gagcacgtta 27960 caagcgtttcttccttccaa gtgctccatg gaacagagac ctgtaatgaa aactaaagta 28020 ggttgtcctagatcagagca tgcaagtgcc caggccctca ggcccaggct gcctgcttgt 28080 ctggctcccacactggcttc ctcacagagg cagtgttccc acctagtagg tctgcacttg 28140 tagtaagtacctattcagta agtgaaggct tatggctcac aaatacctga tgggatttaa 28200 attccaaagagctgtgcagc ttacaaagtt acataaatgc acaggaccac tgacttttta 28260 ttttagcaaagtaagggtga tgcttatctg ttgttgttga ttttttttta actctgtgga 28320 gagagatggagagagagtac accacagtgc acgcatggga gccagaagat aagccaaggg 28380 agccagttttctccttccat cctctgggtc cctgggatca aacgcctcac ggcttcagat 28440 tggcagcaggtgccttcacc tgcccaggga tgctctatga ctccagttgc tttggaacgg 28500 tttttttctaggtagcatag tcagagctgt gagatttggc agactgccac aggggaaagg 28560 acagtgtgtttgtcagaata ctggcggcct ttagaagcga tttccatgaa gctgaagttg 28620 caactgacattttaaaaata attaaggaaa gagagcaacc gaagtctgtc ccgggcggtt 28680 ccaaagaggttgtgtgtctg ctctccagcc atcagcaggg ctgggatatc cgagactaag 28740 tgacaactcaggcaagcctc cagggtcaca caacacagcc cctccactca ggtctcccca 28800 tgctggtagaatgtagcatg caagcctctg gggctgtagg gtctgagtgg gcctttggca 28860 gcctttctctgtggctctcc acacagtaga acgagagatc cggcctgaag gctacacagc 28920 tgtgtctgaggcagagctga gttgctaata tctcttcctg atggccaagg cagggatttt 28980 tacaggcctagaaatctagc cctgcttcgg tagctctggg aggaggtcct gggtgctcca 29040 actgcttggccaggggacag atggagcatt gagcctttca ccaggatctc atggaaagcc 29100 agtgtcctgtcacctgtcac ctgtcacctg tcacctgtca ccattaatag gcacaaagag 29160 tgttgcacagaaaaaaagta ccaacttgtt ttcttttcaa ctgctgggct gggtaatgat 29220 gtaaaaacgacattatccct aataaacgtg atttgcagag atcgttgaca accccagtag 29280 cagagacttgcattagcagg taaacagata agagaaacag ccggcttcac cagctcctgg 29340 cgtggcacgtgtgtctaggt ctggtatgaa ctgaaggttg ggggtggagt gtggagtttt 29400 aaaggcgaatcgggtgatgg agagagtttg tcttaaggtt ggccatccca taaatctact 29460 tctcgatttttaggttgtgg tttcagttgt atacatgttt gttttggtgg tatttttttt 29520 tcttaagggatgggggcgtg cttatggggt gtgtggcttg tcctagtttc tcacctccat 29580 attacctcaccaaaggaggt gggagccctg gtcaggccct agcaccgtcc ctgccaagtg 29640 actaaagagggcagccacat ctgtggcata cagtctatgg gcctgcagcg agtggtagat 29700 tgctcgattatgtcaaggag ttgggatcaa gacaggaact tccgcaggtg gggagagagt 29760 ggcttctgtctggacctgtt cccctagtga gggctgactg gcagctggct ccctaaaaca 29820 cctgaatgtaggtagcaggc acaggtaccc atgtctgtgt gagaatagct tcaggatatg 29880 tgggtaagttagttgaaccc ttgggtgtta aataacctgg atacagtcac cgttatttct 29940 ctttaccatttttttttctt tgccagaaag cactaaagca ttaggactct ggcttcctgc 30000 tcctgaggctggaggagtgt ggcttgtcta accttctcag cagctggcca cgtcacatct 30060 gaaagagctacctgatgctg ttgttgcctc tgtgcgtgtg tgtgtgcgtg tgtgtgtgtg 30120 tgtgtgtgtgtaattcataa gcttgccttc cacctgtccc tcagaggaga cccccccaag 30180 ataaggaataactgaaaggc cagaacctca cagctgagga tcaatcaagt ctcagtgctc 30240 aggcctgggccggggaggag gcatccacat ggactgcgga gagtggctga gggagcctct 30300 gcagggtggcaggttatgct ggaccttaaa gcttggaagg tcagaaggaa gaagaccctc 30360 tactaaggcacaactactag gacctcgctg gatggccggc aggatgtggc atgtggatct 30420 acatgtatggggggggggcg caaagggaca cagctggaag acaggggcaa acatctggaa 30480 ataaatgaaatacccatgca gtctgccaag gggtatagcc tggttaagga attgttttca 30540 tcctgtgggtaacgtgtgac ctgtgcttca gcaagaagac cttgacaagg tctctgagga 30600 cccagccggaatacggccca cagcagccca gccgacacgg ctgtacttgg agcttttaac 30660 aaagacattcatttctcttg cctatggtgt caaaagagag attctcatat gtactgtcca 30720 gtgtggccaaagcttggcca acagaatggg ccgaatctaa ctggctgctg tgctgcctcc 30780 gatcacttgtggggcagtgt gcacacttag tcaccccact ctgccttgcc acctttctcc 30840 tgcgctttgctgtctcctga tcactggccc ctgtccttcc ctcagaggta tttgtgtcct 30900 ggcttcctggcttccttccc tccacccact ctcccttcca attacctctc caagtctttc 30960 tgactttctcctctcacact ctggtgttga ctggggagta aaccagtcct ccagaacaga 31020 acttctctgcaggctccctg aggtcagggg agccatctcc acttgtcact cttgctggaa 31080 gaccacacatggaagaaggg aatcatgtct gtgcaatgag tgcagcgagt aagcccctgc 31140 tggggaagacagcctgatgt cctaggttgc tcagggttac catctgagag gaagcctttg 31200 gcatttcccgtggcttcgga tgacttcttt gcaaaggaat ggagtaaagc ttcctaaata 31260 tgcacagatactcaattctc acagggacga agaagggaca aatttgggag aaaacaagag 31320 cctgccctgtggccgtgaat cagacccaga aagccagaca tgtgaccatg taaacggggc 31380 acatatcggtgttcttgcag tagaaccagc aagattctct cagttggttc cttttctaaa 31440 aacagggtctcatgctgccc aggctagact tggatttgct atataacggc gggtggcctt 31500 gggctcctggtcatccttct gggtgcagag attactgtcc tgcgtcccca tgcatggctg 31560 ctgcaggacactcatctcgg gcttgaacat taggcgagca ctctaccaag gcgagctaac 31620 tcctcccagagatcctgcag ggtttccccc ttgtctgtac gtgttcccaa acccgtgcca 31680 cagctctgaccctgaattgg attagaagag cacatcctga ggttcttcat cttaacttgt 31740 gaccaagcgccagtcctgac gaaaagacca aaacactttc tgttcttctt aaaattaaag 31800 tgtctgaagtagagagaggc tcagccctta taattatgag aagtttccct cgcccaacac 31860 ccatctgcttaggatggctc cccacacctt tcctccttgt cctcttcctc ctcttccttt 31920 tctacatcctaatgtgtaac ccttgtaggg gactttctgc ccccttcttc ctgcgtatac 31980 ccagtgctggccgcagaggc agcccagctc tgttttctca tgatgcagtg attatttttg 32040 gcactgcgcatattttctct aatgttatta ttgcctcctc cacccttctg ggtgccttct 32100 gaggagcactcagtttttgg caattccaca caaaatcaga gggttaattt tagttcagtg 32160 gtgaagacgaggcaaggaga gagggggatg ccttctcctt tcgccccact gcagcatcct 32220 atgcccaccccaaaagggat gcgtttcccc atgcctactc ataaaagagc ttgcttgctt 32280 ccctggctctgtgttagcca ttcatccact gctgggcctg gggttgaggg taccgctctc 32340 cagaggtgacgtccttcggg ggctgcacct cagggctggc atcttaatga cttgacttgg 32400 cgggcttagaacagcctcat tcagaccgag ttcactccct gcgcagttgg cccactcagc 32460 ttctgtccatagagttctct tgttcaagct gcaggaggaa atggagattt ccaagtggga 32520 agcagccttcccaatgcctt aactcttccc tgcagggaag aggagctaca gagagagagc 32580 aaaagaaaacccaagaggca cagctgcagt tccctgggag agggggacag gggcggggtg 32640 gggagcagtgtggctggggg ctgaggctgg agccagcaca gctgggatca ctttccttcc 32700 tggggaggtgggaaggaaga aagtggaggg cgcatttgaa ttgccctaca tcaattagca 32760 gatatttttcagtttgtcca gagctgaggc cctgagaaga acatgcaaaa gtagagaatg 32820 cagtgtctctgctgccacag tccttaaagc agtaggaaca tcacacagga aagccggtaa 32880 gatggggacattctctaata atgaatggcg ttagtaatgt gggcagaagt gccagaggga 32940 gccgggaccaggcatggcaa gaaatataca agtgaagctg attcttcctg acagaggaag 33000 tggtctgatccgttacgtag taagtaccct tgaccaaaca tggcgttggg tagtggacac 33060 actgcacactgtctttggct tcaagatctt aaaggtcctg gaattctttc tgttgaaagt 33120 gtgaggtcatagaccagcag catcagcatc agcatctagg atggtgctag aaatgtagac 33180 cctcatgccccggcccagcc ccgaacttaa aagtacagtg gatcaagttc cctaatgctc 33240 tgtgcactcggaacagtgta ggtgatgtca tagactagat gatagcccag tgtttattca 33300 gagggatggatcacatgctt gtgtgcacgt gggagcaaac acacacacac acttttacct 33360 atgagtgtcactgtattaag aactgttccc gggtagaaac ttcttggtct aatcacgtgc 33420 tgggttagaattcttaggga agattcaaac agtgagcagc attgaggaat ttacgcccca 33480 gaagtcactgaggtggtttt taatgccttc ctctggtact gcttgtgcct cagaggaagg 33540 actctgagggaaccaagggt gggggccttc ccagaagaac atgtctgcat ggggtgaaat 33600 gaaagggagaacagcatgag gctgactctg cagcgtggaa tctctggagc aatgtgaaga 33660 ggtcagaacaggcagggcct tcccggggaa aatggacacc tctggaggtg aacagaggag 33720 ctgtggaacagagatagact aagatggtca agaggaacat tctggaaggc cgtggggagt 33780 gtcagtgcctagagctaaat cttccaggct acgtgggact ggtcagctgt tctccacacc 33840 tcggagccttgcttcatgct agggagttca tgtcacacac cgatcagcct cttcttcctt 33900 ttttctgctgtgcgattttg ctagctctct ctgctgaaaa agaagtgctg tagggacggc 33960 ttgcacatgtgctgtggctt gagccaaatc acagatttgt gttgttgggt gcttgtgggg 34020 tccaaaagaaggtgtctgag aggacacagg agccctaagg agaaacccca gaggccttca 34080 ggcaacagcttaggcatggg gcttacgccc agacccaggg gaaaggcccg aaagaaacgg 34140 accagggagaaagacgcgct caccggagac atccattaca cctgcccacc acagtaagca 34200 tgctgtccccaagtcactct atctctgctc aacccctgtg attctctcca ccagcccttt 34260 ctccatccctccctccctcc tttctttttt tctcccttta acttcttgtc tttagatctc 34320 caaacaaagatgatctcccc cttccctctg gcaagtttgc cccttgaagc aatggcctag 34380 agtagaaggtgatcctgctg tcccctctct tgccacttcc ttgatcaaga gagtgttttc 34440 aatggcttcaaattcagtat tcttagaggg ctataccctc cgtgtgccca gttcagcgaa 34500 gccttctgagctgcaagagg ggcctgttta ttggcatttg gagaaaattg cccaattaca 34560 acccaatgtggcatgtgggc tgcgttgaca cagatgtgag agctaagcat gccaacttcc 34620 tccatctctgggggctgctt cctcagggca cgtgacctgt cgccatactc tttccatgag 34680 atttgaggattaagtcaggc agtaggaatg gataagtgat ccttgttaag tgcaaagcac 34740 tgcccgtggttatttgctag tttcagcagc ggcagcagca gcatttcaat ttgctgatac 34800 taattaaccccttagctaga gtctgcacat tggcagaggt caggcagtaa agaacatcgg 34860 aggccagcaaagaacaacac ttagacagac aaatggccac tgctgcttgt tggcccttca 34920 ttacacgtaaacgtctacaa gtcttctcta gacctccatg tgtgaggaga gggacaacgg 34980 agagagagctagcttagagt gagggaagag gaattgttga cctgcaagat ggccatcatc 35040 ccggcactggcttagagcca aaggcagcct cttcagatgc ttcaaaaaga tctaaggaaa 35100 agaggaaggctgagaggaag gaagcctggg ggcggggcat gtagagccca ggaccaggca 35160 gaacatgagtggttggtttt ccttccttct gcagactccc ctgcctcagg agtgaggcta 35220 cggatgttgccactcaggtg aggggatgta agatggcagg gagttagata catgttacaa 35280 agcagtatgcagtgcagagg cctacatcat ggcactgttc acaacagcta agcgttgggg 35340 accccaactgagcaacatat ggggatcgcc aaatgcattg gggtctctgt tcacacaata 35400 cgctgtgactggcttttagg aatattaaga aagaaatctg agcattatac gtaatgttaa 35460 gtaagaaagtcaaggggaaa agccgtacgt tccgtgagag tcctttctgt aagcatctgt 35520 gtatttcccaggttaccctt gccgagtagg atttgggctg attctcggtt gcatgatgaa 35580 aggcctccttccaagcctag agctgcttgc cagcacactc ctcacgagtc cttgaaaata 35640 catccgaggagttccatcta cttccaccta tccctatttt ctaagcctca gttttcctca 35700 tctctaaaatggacaactgg cagcagctgt tccttcgtgc tgtgaagtga gatttactta 35760 ctcttaaagtgccttataag gtgttgtgtg tgactcaaat gtaaagtagt attcactaat 35820 atgctagtgtttacctattg ccacgggcca ttcagaatgc tgaagcaaaa gccataggcc 35880 gggaaactttcaaacagcag ggagtcattg cttgtgtgtt tgaagtctgg gcagcaaaga 35940 tcaaggtttgagcctgatct gttttgttta ttgaggatcc acattctgct tcacacagtg 36000 gggctggtggaaggtgccag ggatccaact gggccttatc ttacccagag agagggctcc 36060 accctcactttggaggcaag gatttcaaca ttaactttgg agacataaaa ctcagacctg 36120 ggcccttgctagaataaggc taggccaagg acagtttgtc acagctactc ctgtgcgtgg 36180 ccagctttcctagcaggctg gggactccac atgtcctaag gtgatagaag ggtctgggtt 36240 cccagatggactgcttggta attaaatctg ttactgtctt ctgggaggct gcctggggca 36300 ggaggctcgtccgataagca tctccagtcg gcccctgtgc agaattgacc attaaagggg 36360 caagtggagtgagccccaga cattacttac tgtcagctct gaacgtagtc caggcctgct 36420 gctctggggatactgaccct cagagagggt cagcagctgg gggctaaact ccccatgaag 36480 gacggctgggctgaaaggcc attataagga cttctcattg agacggggca tgagagccta 36540 gccctcatttcagccactcc tccctctgct actctgttgc tggcctcccc ttccaggaca 36600 gagaccacactcttcataaa ctgtctgttt gtctgagtgc actgctgcct ctctgcctcg 36660 tccaggtctcagttcttcct gagttcttaa ctccgggtct tccattttga ctgacagctt 36720 ttccttccctttgttttgca tgccctgact gaccactact gccttgggtc agaatgcttc 36780 cagaaagtggctcatcagaa cattgtctcc atagaccacg ttctcgctag cctttagaaa 36840 ttaccctctgagaaattctt gtgagttgtc ttgttctttt gagtgcctcc agttgtggca 36900 aaaaaaaaaaatatatcagt tgagagcaca ttttattctt tccaagaact atgagctgtc 36960 catagcctggcctagtgact aaaagggtgg gtaagttggg gaacatacat agtcagttgg 37020 aatgatgtcattgccattaa atgttgtaac tggtattctt ttgtggttcc aacattaatt 37080 cctaaatcacctaccaaaat gttagagtag cagccgcctc agcaggataa gcctcagcct 37140 tctcctgaagtgactcttgt aatggccatc acctttttgt gatactcggt ataaatctct 37200 atgccattgtttgggtccct tcctgtagct atagcatctg tagagcaatg gccccaccag 37260 ccctaacagtatctgttcag cctatgatag tgactttaaa tctgcttgac atgatggcca 37320 tgacaccgtgtgagggagag ggggagtgca tggtcagatc tcagaggtat ccgaggactt 37380 cctgctttttgtgatgtata ataatgggtc ctgatgtctg tatcaataag aacgcaagtg 37440 attttgatatgagccaacat tgaaaatggc tgttttgcta aaatgacatc agtaacaata 37500 attccaatgtaaacatgggc caaaaaccaa aaacactcac tgaggaaaag ccctgcgccc 37560 caagctccataaacgcaggt tttctttatt cctgagtgtt tgagaaaagg ggtaattgta 37620 tttccaacacatccttaatt ccagattaca tacatagtac acccccaaaa tcaacaaaag 37680 ggccctttaaaatcagacag ctttgtccag gtgtggtggc acacagacct ttaattccag 37740 ctcggaggcaaagccaggtg gatctctgat tttgaggcta gcctggtcta cgaagcaagt 37800 tccaggatggccaggactat attacagaga aactctgtct cagaaagaaa aaaaaattga 37860 cagctgtgtaacaatggtta gccctgggca cataagaaca gaattgggca ggagtcatgg 37920 tgtcctcagataaatcaaat ctaaggtcag tccgagctgg gaccccagga tccatttttt 37980 gggggggtcgagacagggtt tctctgtgta gccctggctg tcctggaact cactctctag 38040 accaggctggctggccttga actcagaaat ccacctgcct ctgcgtccca agtgctggga 38100 ttaaaggcatgtgccaccac tgcccggccc caggatctac ttttaaggct tttccagtga 38160 gcaatcaagatcaagaactc tgcagaggca tgggttctgc tgtgatttca tcagttgcgc 38220 aaacaactgctaagcttggg gtccagggac tcttgatttt ctccgggacc ctgagtaatt 38280 tttctttttaaaatatttat gtatttttat gttggatttt gtgttggctg tttcgctggc 38340 ctctgtgaccatgtaccatg tccacgcagc acccacaaag gaagaagata tcagaatcct 38400 taggactggagttacagaca gttgtgagcc accatgtggg tgctgggaat tgaaccccag 38460 gtatggaagaacagtgtgtg ctcttaacca ctgagccatt tctagcctga agctgctatt 38520 tctttcaccaggcagctgtt gtctggcagc tccacaagct cactgaagag cccacctcct 38580 tcctgcttgccttcacagtg ccctgtgatt tagcgtacgt ttagatccaa ccaacaggtt 38640 ggcccaagctggtttagtga gcctcgcttg acctctcagc cacttaacct tatacggtag 38700 cagacatctgacttagatac ctgatgactg cagtcacagt aaaagttgag tctgctggag 38760 acagctaggcttggacactc gcagatgaga aacaaggatt gggccgagag taggtcactg 38820 tgaatgagagcatcgggacc cactgccaca cttacagtat cacgtgctct ggccaagctt 38880 tgcctgggtgagttttacct catagtctag gcttctggat cctttgattc tactaattag 38940 atctaaaatatttggaaaga aattatgaat gtggtgaata tatacagtct ttttttctta 39000 ttatcggcccctaaataata cagcataata gctacttata gtgctcacat tctaggacgc 39060 attgtaagtaatcaagtggt ttaaagtata gatgcagatg tgtgcaggtt ccacacaaac 39120 actaccccttaaggactgga gcatctttga ctttgtcttt gaggggtagc cagtcgtagc 39180 tgagcgaggaacggttaccc ctggattgat gcttctggac agtcagttct gttttacctt 39240 tagcatgctgcttagaagag cttacctcct ttacctggga ttaaatcttt cagaatcatg 39300 tttgtttcccctcagggaga gctactgtaa atgaatgtgg aaaagaaaat gctcccaccc 39360 ctagctgcagtgtggtagga aggactgtgc agtgtggtag ggaggactgt gcagtttggt 39420 aggaaggactgtgcagtgtg gtagggagga ctgtgcagtg tggtagggag gactgtgcag 39480 tgtggtagggagcgtgcaag gctacttgat gttggacttg atcaaacaac acctcactgt 39540 aagtcagctctacagcaacg ggggctgtgt gcgtcctgcc ttcctgctcc tcccagtctg 39600 aactccaatactggccttac tccttgggtc ctaggatgtc cctaaggtta ttgggtgctt 39660 ttccaaagacagagctgtcc aagccccaag gactgctttg cactgtgtct ttctaagtct 39720 caagcctcctccctttccca tggtgtggat ggggatctga gttgtaggaa ggtctgtaac 39780 aacactggacatgtcagaac gatccaagag tttccttgct gcatggggag aaattggggt 39840 gttgggacccagagaagcta ggattgagac aagtgattta gtctctgtag ctcctgtctt 39900 aaagagttattatgaaactc aactgcagta aaatatgtaa aatatgtacc taagccgtta 39960 taagggcaacattgttatca ctcgcagtta gcagaatcct gcctcactgc ctttgctatt 40020 agcaggtttgtctctgatcc tctagctaag gactgcaggt gtaggtgacc ccctaactga 40080 ctgcttacttctagagaagg aggcttgtgc tgtgaccaca ggggccagca tggtccccgc 40140 aggccaactttggtgtgtgt aaggacaggc agccagaacc acatttaggt ggtgtgtttg 40200 gaacaccacatatgatacag gccatctcgt gggaacagga gagggatggg aaccaaggat 40260 gggaaaggaacttaggaaag aaaaccaacg tggagtaagg aagtaccaag tctctccgaa 40320 tcaagtatttagggtaggat atctttttaa gagaagccaa tggattctaa tatgtaaatt 40380 gtgggacacattcaataagg ataagcaaaa tgtggtcatt gtaggaccca tcttggaaga 40440 aaagtgttccaggaaatgaa gcacattgtc ggcttaggaa gatgcccaga tataatatgc 40500 agagccagtcaagatcagcc cgtgtgccat agcctggtcc cctcagacag cctgcttagc 40560 gaggacggacaggtgccatg ccagagtcca gatgcagctg attctcaggg catcgatgcc 40620 cagggtagccagacaatgga tcttcaatct gccccaatag ttttgattgc cagagaacct 40680 tctagggattctgcacccat tttatcctga taagacatgg agttttcttc ctactttaca 40740 gatgcaggaaccagagccca gagaacgtgt tttcttcccc agattgagat gggtagatag 40800 cactgtggcttcgtacaagc tcgggcttct gttatctgtg ccttcatgat aattattctt 40860 tttcttcagggttgctaatt aaatgatttc agtgttacag atttttgtct atttttccaa 40920 gagtcaacattagacatctc tggactatgt caagattaac taggcaatct aattaaaatc 40980 aagctagttctacagtggaa ctggaaaaaa aaggtagcta aagggagtgt acaacatttt 41040 aaaccagggacctgcctcag ggtctcggcc atctaaatgt taaaatgttg aagttgcctc 41100 tttccagctaaggaaaggta ctgcctctta tgcgagtggg aggaggtaat gttttatcac 41160 tagctctgccactaattagg taattgctct aagcacagaa cttaaccaaa ttgggtctca 41220 gtttccaagtctctgaaatg gagacaatgg ttgcaaggat aaaattagtc agcctgtctg 41280 ctccctgactggaagggcct atgtagctcc tggttgtaag accttgggaa aacggcatgg 41340 tatgttctgggcctcagtgt tcctatctgt aaattgcaca atgtctaccg agccgtgtca 41400 gtaagaagagtataacgggg tgatatgtag ttgtcggcgc agtgactgaa cgtgtctgta 41460 tcagtaagtgtttatgtagc tgaaggagct tagccaaacc cagagctctt atgccaaaga 41520 gaacccagactttagctagc ctgttcccca caactcagcc acgggggtag ggggcgcgga 41580 cgggagagcttgttcttggt atcgttgctg atacacggcc tgtggtgact gcttcacggc 41640 atagctgctctggatgttaa caacgacggg atcaggcgct gaccctgctg ctgtccggaa 41700 gcgtgagggctggtgctgag gaggggaatt caggatctcc tacttggacc tcaggagcca 41760 gagctgtggtactccagtgc agccattcct cctgtgagcc cttaaggtat cccaccctaa 41820 ggagctcaggattgagatat aaaatccagg gaccaataat ggcccttaaa gtctggtaga 41880 agatgcaaattctcccaggg gtcaggttct gaggggtgag aagggtggga tagaaataga 41940 gaggtgtggggtttctgaga gctgaaaggc agggaagggg gagaaaggga gacaaggaaa 42000 gccaaggggagaggggacaa gaaaacccat ttccctcttt cacaacttct cacaaggttc 42060 tgcctgaccatccatgttat gtggctcttc ctgcagtctg gtatccaatg gctaatccat 42120 ctgggggcctagatggcctg caaatgaagt gagctctgac gtcgaaaacg tcgaaacagg 42180 gcctctgcctcaaatccgca cggggatgag aggcatgcca gcattccagg aatccccaag 42240 tagtgatgttctgtccagat aacgacatgc tcaaagacag gcagaaagga gagcaacccc 42300 taggactggcaacctcagag ggtaaggtgg catgagccag cctggagcta attggaagaa 42360 ggccttgaaagccacaaagc acactggaca tctacagaag caaataccaa gttagtttct 42420 ttattaaacaagcaatatat gttatttata gaaaacacag gaaaatatcg ataaccactt 42480 ggtaggccagggagggcgag ctccctaact aaccccatta ctctgcaact cttactaatg 42540 gctaagtgcctagactctgg ggttgccctg ccaggggcag agttcaccta ccagctggca 42600 gtcacgggtgaattacttag cctccgtggg cctgttttct tatctgtata ttggagatgc 42660 taacagcagctactctcaca acaatttgtg aaatttaaag atgctaacac tgtactgtct 42720 gaaagagtagctgaactgta tcaaaaaacc tgtcaccatg acgctgtgac catcgtaaaa 42780 atgtttgctacttaactgca ctccctgtgt agcacacagg aagtgctgtg tgggacctgc 42840 acagtgttttgaggacatga ttgccctctg ttgcggatag gttgtctttt catggacaga 42900 ttgttgctaatgtttcttta tagtggaatg tgcccaggac taaaagtttc acataaataa 42960 atggtcacagtatgtcctca cagttactgg ttactgatgc gacacttagg cagcttcatg 43020 gtagaatctgacgagttagc aggcagatac tctgactttt aaacttaccc gtgttagtac 43080 gtgatatggactttgtacga agaccgtgtt tctttaggat ctctggaaag aggcaggttt 43140 gggtgtcagtttgtcctttc cttcccattc tgcaacaaag aagagtcagt ctggcacctc 43200 aggctggcaaggatggcacc cactgcagct accacccttg gaggtctttg cttctggatt 43260 gcaaatggaggcgtgttgtc cgcctcatgt tctcttggcc tttactgatg tctccagact 43320 ctaacctgtcgtctctcaga tcagaaacag ggtcttaggt aagccagggc tggtctgacc 43380 gtagcttcttcgcccttctc tttccattgg tgccctttga ccctgtcctc aaactttgtt 43440 cattagtttaattaaatctt tgctaacgct acccacgtga agcccagttc tggctcctgc 43500 aagaatacagaagaaagcaa tttgagaaga caccaatgcg caaaagcaga gtcaatacca 43560 aaaggtggcttgctcatagc tcccctgggc tgagccagat gggttcagtg ggagaattga 43620 ctcactgtgggggtgagtgg gtcactaccg agagtgtgaa tggatgacgt ccacattcca 43680 ggactaacccctcgtttctt catgtaggag cagctcagag ctgaggaagg agaaatcccg 43740 tgatgccgcgaggtgccggc gcagcaagga gacggaggtc ttctatgagt tggctcatga 43800 gttgcccctgcctcacagtg tgagctccca cctggacaaa gcctccatca tgcgcctggc 43860 catcagcttccttcggacac ataagctcct gtcctcaggt aaggcttgac aggtcctgcc 43920 ccaagctggcatctacctag gcctcgctcc aagacacatc tacaaatatc cactcacaga 43980 agctggcacatggcctttag tgttacattt atttagttgc gtgtgagggt atgcatgtgg 44040 gtcagaggacagcctttggg agtccattct gttctcttct tccatcatct gggatctggg 44100 acttgaacttgggtcctcag gcttagcagc aaatgcctct agccactgga ccttcttgct 44160 ggccctgttccttcatttta gcatctcccc tctggcaatg atcttctcat gagttcaccc 44220 agggaagagaccaaggacag actcaagtga gagtgtgagg tgctcccaga gagtgtgagg 44280 tgctcccagagagtgtgagg tgctccaagg ggttggagag ccgagagcag cttctcctgg 44340 aagcccatccagtacctctg gacctctggc gagagtcccg ctccacactg tgttgactct 44400 gcaggaagccttttatcctt gtcttccagc tacatctcta ggacatcaga aatggtgatg 44460 tcccttgtgatctatctctc agaaccttgg tttccttgcc tacaaactgg aattagccag 44520 gcatactgcctgggaggata ggggtaggaa atgggggggg gggattatta gggcactata 44580 ggaatgagtggagacagcgg ctcagctgta ttcgttcttg ctgggctagc ccccgccata 44640 gaggacagcctcgggcacct ctccctgctc agccgatgcg ttcttctttc ccgcatatct 44700 cttcaccaacaaacagttca taacgaatgc tttctttcct ttgtcagagt tacatccctc 44760 aaaaatcatttcctgttagg cctcaccagg aagaggcagc ctggggtttc cactttcaca 44820 tcctatgtgcagtcttgtca gacttatcag ttctgtaagg aaactgggca gcatatagct 44880 gccaggctggcactacagca gggcagtgtc cgaggcatga gcaagggagg caggcaggca 44940 agggggaaagagatcccgtg gctcattttg agttttcctg agtgagtgtg tcactctgga 45000 gatgactccttacatggcta ttctgggaaa gagccccctg cacagagggg tccagaatga 45060 ggcggggaagccagactagc ctgtgctatt ctgggcccct gtgcacagga aggatatatg 45120 ggaaagaccttcggaggtta gaatggctgc tcatcccatc gtcctcctct aacccccagg 45180 ctggaggctaagcctgggct gcaaggctga ggtgaccgtg ctgttacaga aatgagcaga 45240 gagtggagaaagcaagggcg gagccgctgc acacacagca gggcaacagc aattactcag 45300 atttagacggtgaaaatggt tgagggaagc tcaggctaag gacttgtaaa gcctggactg 45360 ctaaataaaaaggcagactc ggaggtgtct cacccatgcc ccatgcatgc cttcatttta 45420 cagaggattgtcctcttgga gaaatgagga cgacagttcg gtgatttgta ggattttgca 45480 aagcctgtcaggcaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaagaaa tgtagataag 45540 gggcagggagccaatgtcca agtgaagcag ctagagcctg accaggacta gccaggagca 45600 gtgggtggccaggaggttct gagagctgtg tcttgctgcc gtagcaggga cacattgtct 45660 gtgctcgcccacacagaagc ctgtgtgtct tcctcgatgg gtcgaggttg atttgcagag 45720 ggcttggctagggttggatc ttccgagctt atctgccctc atgtgtcctg gtgcaacccc 45780 tcccgcactccacgtactac acaaagccac agatacaaga gcagacacca cacggagcag 45840 acatctcaggagctctgagc cttgagaaca aggactgcct actctctaga cagcataagc 45900 acggacagaccagaaccctt ggcgcgtcag ctatggggct cccaggcctg aagaaagaaa 45960 agttagagattgataaacaa gttttggtca tctggtcctg gtgaccttaa agaagtgctc 46020 ctgagtccagccacggaagg agatgtggct tagttctcct tctctgccat ttctccaggc 46080 tcctaccaggcactctcggg actggttatt tccagaaatg gaatgtaaaa tgagcctttt 46140 cctccccacccaccctttgt tttagtgtgt gcatgcgtgc tctggagagg ttagggaaga 46200 gcgtcgaagtcttgcttaaa gacttcaacc tcccttcttt tagacaggac ctctcgctcg 46260 actcgaagctcacgatttta gctaggttgg ctggctggca aactcacagg atcctgactg 46320 tgcaggtcaacattggggtt ccgggcacac acagccaacc tgtcaatgcc gaggactcga 46380 actcacatcttcatgcctgg gcagccagtg ctcttatgca cttagccacc caagtggctc 46440 attgttttaaattttcacct attatatgca tgtgtttgtg gaggggagga aaggacaact 46500 tttgggagttgattctccct ccccaccatg gatagggttc caaccaagtt gtcaggtctg 46560 aatagaaggcctttttacct gctaagccat cgtttcaacc ctgaaccata ggtctttatg 46620 ttttgtttttgttggttagt tggtttgggg gttgtgtttg tttgtttgtt tgtttgttgt 46680 ttgttttttgagacagggtt tctctgtata gccctggctg tcctggaact cactctgtag 46740 accaggctggcctcgaactc agaaatctgc ctgtctctgc ctcgcaagtg ctgtgggttt 46800 tttgtttgtttgtttgtttg tttgttttat gtgacaaaaa gtttagagga tctttgagca 46860 gatatcctcctgcactttgc ttattggtgt tgctgccatc tctctcagaa acattgtaca 46920 cagctctatctcattggacc gcagagtcca tgaaacattg ttggatgata tgaaagtcta 46980 gcctgttgtacaagttatag ctttgaagta agtctaacaa aagaaacgat gtaagagaaa 47040 aatcagagcgaactctaatg tctttggacc caccttttag cagttacgtg ttacagtgtt 47100 acaacatatactttcccaac tcaaaacaaa ctacagactc attacttagg caagtggagt 47160 tttgtatacctcagagttca aacgcctaaa aaataccagg cttagcgtta gggccagttt 47220 cttctttacttagcagcaca cttcctttga ttttcacagt aggctgcagt gtgtgggaat 47280 gttggggaggaagcctccgc gctgagaact ccaggctgag tcgggccaca gttgagattc 47340 atatcacaggaaacaaaccg aaacaatagc tttacgatac ttgcttccac actggcccag 47400 gaggacagaacacactgtgg cgggaacatg ggtggaaata tcacttgatt gtcttaaatc 47460 cagatgaaccctgcgctctg gggctgaagt ggagtcgctt ctgcgtccca agagctttag 47520 accgcagtaaatgtatagaa tgtgcattcg ccccaattct gatttgaggc ttcccagact 47580 catatgtaaaaaaatcaaat tctcattact gcagagttgg agatcagcac aaagccaggt 47640 ttctagacataaatgtcaag tttatttttg attattttga tttgaatttg tttatgtttt 47700 attcctggcatttgcctagt gaagtcacac agtctgctca ggatatgatt ctccgatccc 47760 tgagacattaaaatccagga catggtttta aagctttcac catgacttct caggaaaagt 47820 gggacaaaggggacagaatt acagcagcag atgtgatttc tgtgccctcc tatgccttgt 47880 ggtaagacctgttttccctg gttttcagcc caattgtttt actgtcccac ctcccccggc 47940 cccacctatactcaaaatca aggccttttc tgtcctgttt ggaaggaggc cagtaagatg 48000 attcatgccaggatgttact ggctgagagc agccagcggt cccttcaaga aagtctaacc 48060 ttgcttatagcattctctta aagcaaagag tctggccagt cagcgacagt cactgactgt 48120 agcgccccatagcattttat gaaggctagc gcagcaagca agggtggggg agcaggtgtg 48180 aaaagaacaaaataaaaatc tccaatgctg gacttgtggg gcacaccagg agagcagcag 48240 caaggccagctgagatctat cactctgcag aaagtgtgag atagccccag cctgctcaca 48300 gtgcggcataaggcacagta agtggcccac actctttatg tttgccgtca gtatgcccgg 48360 aagacgcgtgcacagccttt gaaaggaaag accctgcgga gataactaag tagcaagcac 48420 cagggaagtaggaaacctgt atcggagctt gttaggaaca aggagtttct tgaagatgga 48480 aacatctagaaggatcatcc gggtgaagta agaaagcagc agccttacgg ctggcacagc 48540 caggcctcaaagacccagtt agaagccacc tgctctgcca cctgctagtt cacacaaggc 48600 aagtggctctaccatactgg tgtgccaccc aacatgggcg gtgctgccta aaggaaatga 48660 gcagtgctccggaaaaggcc ctccacagcc ttctcagcgg cacatatcct ggcggtggga 48720 gccatcaaagcctgtttact ggggctattt ttagcattaa agaatttcgt ggtccttctc 48780 aaaggagacagttcgtctat accagttctt tgagattcga accctgacag attctgggaa 48840 gcaaatggccaggatgtaga acctgagcta tttagaccac ccagcccagt tccttagcaa 48900 gcacctactttattttgtac caatggtttg ctctccgttg ttatcagcat ccccaggagg 48960 ggcttaggctcttcgacaga tgtcttcctg gcagtttgtt ggttcctgaa ttgcaccctt 49020 ccttgcagtatccccagctc tccctgagac aggactgagt gtgtaatgag tgctgtgagc 49080 cagggaagccatggaggaaa agccttagta actgcaggga gggagggagg tctggtgtgc 49140 gcagccgccaggcatagcag tttttagcag aattgtgaca ggaggctcag ggctctgggt 49200 gcagcagggggatgtctgcc tccctcttgg ctgggagtga cctagccaag ttccttcaga 49260 gactcccaggaggacaagca ggtgctaaaa gagcaaatag ttccactgaa ggaaggggcc 49320 acacccaagctgggctgctc tagggtcgca gggaaggggt gggggagggt gctattggcc 49380 attgtgacttcagtctcaag atgttccatg tctgtggccc cagacaccct tctccctcct 49440 ctctaaagggcagtccacct gccactgtag ccaatttcgc cacctcctgg aagtaagcgt 49500 gctggacagttcggaaaggc cgcttggctg tgccgggcct gttaaaaaca ggaaacttta 49560 agcagaactattttctctgg gtctagttaa ccccgatagg ttgtcttggg attactccag 49620 attttgaagtcagtgttgcc actgagatca aagaagctga agtgaaaata aattctcagt 49680 aggcctcagcactagcctct gtctgtctgg agaaagtagc cacctcgccc tataacccaa 49740 atgcagctgaaaccttctcc gggcatgttt ccggggtcag gcaccctttg cccagactgg 49800 ctggttttcctgacgtgggg gatagtcttc agcacgtggt ctctggagcg acagctttga 49860 caccctctgaacactttttg ttgatgttgt tgttgttgtt aaaggaagaa aaggcacttt 49920 ttcagcttccctgaattagg aaggaagcct gggaggaggt agaaccttcc agcaccaccc 49980 tgggtggggtgcggcctcct cgtactagcc aggtcttggg ctctgagctc agcttaaatt 50040 ttcagcagagggttccacgt ttttatttta ctttgcacaa atcccgcaag ttgcatagca 50100 gtcctgggcccgccagaggt cctggcccac ccactcagcc ttggctagac ttgaactcac 50160 tatgtagaccaagatggcct ggaattcaca gagatctacc tgcctctgcc tcctgatagc 50220 tgggattaaagacctgctct aacacacctg gttaaatcca gatttctaaa gcacacacat 50280 atttgacattaaataatgaa caagaagagg gcatagcctg tggtctgagg ataacagcca 50340 ggagccggaacaagagctga gcttagattg cagaggtgga cttggtagtc caggacacac 50400 agagtgcatggttgggggta gagttccccc acaacgcccc ctagtgtctg cctcttgtcc 50460 ctcacggctttgtgcctcta agttccattc tctttcgact attctatgtg ctatctatcc 50520 ccggacttatgtccccaaag tggtgctctg agaagccacc tctctgcccc ttgactggaa 50580 gagaagctttgggacactgg gctcccttat tgtcccagtc tctgatattg ggccatggat 50640 cttctgcctctagctggcct cttgtctgtc ctgggaggaa ggctgtctgt gtgtcctgca 50700 gtggtggcccaatcctgtcc agttgcctga cagacctctc tttccatact catgtgaact 50760 cacattccaggtgaattagc aaattgctct ttctaactct atgaagaatg gagctggaat 50820 tttgctggggattgtgttga atctgtagaa tgctttcggc aagatggcca ttctcactat 50880 cttaatcctgccaatccatg agtgtgggag atatttcatg tttcctctcc atcctatgtc 50940 ttcttttttagagtcatctc tcctctgact gctggagctt gtgctctctg taccccttct 51000 ttggtaccccatggtaatgt gcgtgagggc ttatttagct ttgtgaggtt gtgagccacg 51060 aactcgccaccttggctctg atttgagata gtaatggtgt cttagaggag ggagcagatg 51120 aggtagagcttgtagtcgtt gatatcactt taagcagtta atttacttaa ccttacaatt 51180 catgaagatgggaatcgcta tcctggtttg ctggtggagg actccagcgt tcaatcgtta 51240 cccaaagtcataagcaagtg ggaagcagat gtaggaatag atacaacatt tgactccgaa 51300 gcttgtgaggtggttgaacg tggccctgca cttagctcct ggggcttcct aacattctag 51360 acatcatagcctttggaaaa atggcttgac tcagaagtct tgcactataa aatgagactg 51420 caaatggtacatgccttgtg cgttattgaa aagccaatgt agagtgttta atgctgggtc 51480 tgtctgtggttgatgttcca cttacgttag cagctaaaat aactgctgct gctgctgcga 51540 ggtctagcattctatctgta gcctctaccc ccagccttcc tattggtaca gcaaattctg 51600 ctaccacagaaaccacgctg tcccacagtc attgtcaatg tggcctgggt gttccaccag 51660 gcatgtggcaaatgttagat ggttggtagt gtgcctttcc ctgtgccctg gagccatgcc 51720 tgtcccctcggtcatgtctg ttttaacact cgtgcccctg ttgttcattc ccctctctct 51780 ccagtctgctctgaaaatga atctgaagct gaggccgacc agcaaatgga taacttgtac 51840 ctgaaagccttggagggttt cattgctgtg gtgacccaag acggtgacat gatctttctg 51900 tcggaaaacatcagcaagtt catgggactt actcaggtga caccctctgc ctcgttcagt 51960 aggaaaaacatgtctttatt tggggataga cactaacggg gggtcctagg catagtctta 52020 cttgacttttccttatgcat tcccatatga tgatgacagt ccttaggact tcccaatgtc 52080 atggggcttgacattccttg tggctgccct gacaggtctc ttctagctag attaacttgg 52140 caaaagtataaatcaagccc ttgttgccat caacattgct ctgatacgtc tgtaagtcca 52200 tagacccaatattgactgga gactattgat aaccactcag ttcatccccc tgcctgtctc 52260 tgaatgcagacattatccta gcttcctctt ggagtccgaa tgacttcatc actaggagta 52320 acagcatctggccttgcttt tgaaacaggt agaactaaca ggacacagca tctttgactt 52380 cactcatccttgcgaccatg aggagatccg tgagaacctg actctcaaaa acggtaaagt 52440 gttcttctttgtttgcattc ttctcatgac ccccaaagcc tgcacaaata gcccaaatgg 52500 attatgttccatagatacag ttggactagc ttctgggtga gtatgcagct gttgagatga 52560 ggcccagcacatagaatagc tcctaatggc ccatccatga tgcctgatgt cacactacga 52620 ggtcagggtgccatctctag gacatttcat catcacctga gatcaatcat ctccgccaag 52680 cgacaccacccaaaccaata gcttcatcta tgcctgatta tttatgggag ctacaggtgc 52740 cttttgtcgtgtatcaagcc acccaacaca caggcttaag caatctccca catttctggg 52800 gattctgtgggcaggtctct agctcaatgt gacatcagct ggattgcagc catcaggggc 52860 taaactaggttgaagtgatt aagatggctc acctggtgtg ctgctgggac agtgaccaca 52920 gctctcagtctatggagcct cccggtagcc ttcttcatat gaagactcca gagcaaccat 52980 cttgtatgacagctcaggcc atgcatcttc cttctcccct gtaaagtgca gctgcagaag 53040 cctacagaccatcctaggcc ctagccctga aataggcatg tcaccacttc taaccctatt 53100 cggctggcgagggaaaggtt caggcttgcc ctgtgtctca ggtccgtgtc aagaggcatg 53160 gcccttaggattaccactga agagcagcta tgacgggaat gtggagattt tcagaagaac 53220 taaaactttggtgctggaga gctggctctg tggttaagaa tactggctat tcttctgagg 53280 acctaagttgacctcccagg ttctttgact ggtggtaggt cacaactcca gcttcaggga 53340 gatccagggtcctcccctaa cctctgcagg tatctgcaca aatatgccca taaacacaaa 53400 ggtacaaacttcagcaaatc tttccaaacc acacgaaagg agccatcgct gtaggcagca 53460 gtggcttccaatgactggtg agatcctctt gaacttgaga atctatcacc agtctggaca 53520 gactgcccacatcaacatct tgtaaatctg tcacctgtca ctactatggg gtgtgtgctc 53580 atccaacctgaatagcaaag gcagtgatgc cacgcctgcg aggcgtgttc tcgttgcctt 53640 gtgtgctgcaaaaggggagg ctttgttggc tcacctctcc tgttggtaca gagtactgta 53700 atccacactcagacttacaa agctttgtaa aattataacc acctcccttt ccaatgctgc 53760 cccagcctctcatctgcatt gctgcttcct atccaaagac cttggtcaac tggcttccag 53820 agtacataggctggggacca ccatgagttt atttgtcttg cctgtggtgc caagaaaccc 53880 taggctaagccacccaacac atagaatgat tggttcttga tggtgaaggg gctattgatc 53940 acagacctgcgccaccatac ttggctttca gaagcacttc ctaaatcggt gtctctcgtt 54000 gcttctcttctagagaacac tgctccctgg gttctttgtc tcaactcaac tgctgaccaa 54060 aacctttgtttgagcatctg tgagctgaac taagaatctc actcctgtgc tgttcaaagt 54120 ctttcttttcaaatcgatgg caaaagatga ggccaaatct aggatctttt tgcttttagg 54180 tttctctgactcatagctga gtgtcctcca cttatctgag gaaaactcag gtctttagat 54240 tcatgaatggggatttgaga tcaagcagac tcagccaatc agcaagtcct tgctgaaggt 54300 ttcaggctgcatagactcat ggtctaaaag caaggggtcc gtgtggattc ctggaagaag 54360 aaaaggttgggagacgggct gtggaatgct ttgatggtga gtggaaagcc tcaaactggg 54420 tcctgcaaacagtcaaaggc tgttgaagat gtttaagctg aggtgatact atcagatgtc 54480 agaatggggagactccagtg attagcaagt taggagccca caattacaac ccagactatt 54540 aatcacaaatgaatattgag ctccctcatt cctaaatcct ggggggggct ctctcagtac 54600 agttataactcatcatatac aaactccacc taataactaa aattataaga agcctaatta 54660 tgggagagacagtttataag aacaacattc aagctgtcac ttaatgggtt ctgcagaaga 54720 tgtttaaatccagttcaagg atgccagttc ttttgataca cagctgtttt agactgtgtc 54780 ggtcaaatcaatgttcctta gaggtgtggg tgcagaggtt ggcagcttct cttagagcag 54840 tgtgactacttgttggaaag ttctgaattg ctctgtgatg ctgggcctgt ggaggccaag 54900 gatgaagatggcggtgtagc cacagtttag acaccgttgg actcatgttt tctgttctgc 54960 ataacaagaagcagaaaacc tggacaggct ttggagattt gtactaaagg aaagaagccg 55020 tatgttccttgtctcggtga tttatttaat cctaaatgaa aggtcatcta attgatcttt 55080 atcaagaacatcgttaagat agacttgtct acccagttcc agttaaaaac aaggggtgtg 55140 tgtgtgtgtgtgtgtgtgtg tgtgtgtgtg agtgtgtatg tgtgagtgtg tgagtgtgtg 55200 tgtgtgttcattcccctccc ctcccctcct tgacatgtgt caccttagga ggaaactgat 55260 gaatggtacattatccaaag catagtcatt cgaggattca ggtaggatat tggttgctca 55320 cattcctgagagacaggcta gcgagagcag agaggagtgt ccaaggataa ttctaactag 55380 acagacttatgtccttggag caaccatggt cactctcagg tttcctgcta gagacccaag 55440 gcctgactggggagagacat ctaaaactgt ggtgtagcct cacagctctc aggtgacaaa 55500 caggtacagtgtgtagcaaa catggcacag ctcctgcctt ccaggccttc ctgctcctgg 55560 gaatgcagtcacacaggagc ttctactcag ccatcctcca agaaatctaa cttctgcagg 55620 ggagcagctttctcccagga gtccctccac tgtgagtctc caccaacctg gtgttagctc 55680 atagtccctgggtgggactg gccatcacta gaagttttgt aaaaaacctt ccttgacata 55740 tttctgtcctctagttctga ccacaattga gccaggactc aaggcacagc aacaaaggga 55800 caccattagtatttaagcca tgagggcctc tctgcccatg gcaggctaca cgcactctac 55860 tacaaaccacaggagacaca aatacaagtt gcccagaaac attgtaattt ggattaattt 55920 aactggctcccacgcccttc cccactcagg ctctggtttt gggaagaaga gcaaagacgt 55980 gtccaccgagcgtgacttct tcatgaggat gaagtgcacg gtcaccaaca gaggccggac 56040 tgtcaacctcaagtcggcca cctggaaggt aggattcgtg gagtctcaag aaagagccag 56100 gagcaggaggtgcctgaggc ctctccctct tctcggccgt ctcggccttg tcttacttct 56160 gtgctttgaccccaggtcct gcactgcacc gggcaagtga gagtctacaa caactgcccc 56220 cctcacagtagcctctgtgg ctccaaggag cccctgctgt cctgccttat catcatgtgt 56280 gagccaatccagcacccatc ccacatggac atccccctgg acagcaagac tttcctgagc 56340 cgccacagcatggacatgaa gttcacctac tgtgacgaca ggtggggtgt tgggacaggg 56400 tgggtcttaccagtgtgcat ctgtgagagt gtgacagcgc agggacggga ctaggacatg 56460 gtgtgggactgctggctgca agtttgtaga aggtagcctc cttccatgtg aagactttaa 56520 aatgaagagagctaggttag actctaaccc tcagttccaa agcaactgga cgttctcttg 56580 ggagtggggggcacagaaac aataaagact gtggacttgg actagagaac ctagcagagt 56640 catctgtggtcagtgtaggc tgctgttctt accttcatta aaggggagac cacagagggt 56700 gcacaggaaggcatgctgtg tggttgtcag tacatgcaag gtttgtatag actacaccac 56760 agttcctatccacttgtgct gctgctgttg ggatggagat gtggaggaca acggtgagaa 56820 cgagttgtgatgtgcagtgg cttgcaccag ggtgagggaa gccaggtgag aggctgcact 56880 gggcgtgcatgcctagcctg agtgaaaggc atcactcact gtgcctgact acttcaccca 56940 tcgtaagctcagctctgccg tgtctctctg agcagaagat agatcggagg tacgccctct 57000 gcagttttcagagagacccc gaaagtcccg gtgccagatc catgacaccg gctttgaggt 57060 gcagtggcacttgggatgct tgtgcagaaa cccaggagtg gtcagggatg tgggtgacag 57120 gagggagttgctctgaagca ggaaaaccaa accctcacct gccatctcct gaaagcagaa 57180 aagagactgtaaaaaggagc tggcaggtga gggaactatc tcccagaaag gttcatttgc 57240 tgttaatttccattcattat tgtggtttga gtgggtttca ggtaaaaggg ctagccttgg 57300 gttaagggcagaaggggaca gtcacaagaa atgggcagcc taaagggaca aagatgccat 57360 gtgcacgcacacagaggtac catggtgacg tccttaggtt ggctcagtac ggcgggtggg 57420 gtggggttcatactcacagg aaggatcctg ggatttagag atgtggctcg tcgtgcacag 57480 ggagatgccgcttagggtgg cttaggacac acagtatttt cagcatactc ttgccttcca 57540 aggaagctgattgcatggcc ggcccaacgt gaattctgtt ctgctaggga gccgactgca 57600 gagggatgcaaacacagagt gccccacgga gcgtttaacc gattagcaga ttagttaacc 57660 agggtacagataggacagtt agtgaagggg ctattttaag tgtttaatcc cttggtgtta 57720 tttctcgattgctttgggtt gggggagagg cttgttctgt ttgcatgtgt tgagatgtgg 57780 agcagtgtgtgtgtgtgtgt gtgtgtgtgt gtgtgtgtgt aagaggggaa gggagagaga 57840 gagagagagagagagagaga gagagagaga atacatacac taacatgacc ttctgagaga 57900 tgtgctctaaatccagagcc tacatgtgag agccagtgta gagcagggct cccccatttt 57960 actcagtgaattatttggtt tggtgacatc tgacacaaag gggattgtgt cccctaacag 58020 aagcagggaaagaatctgaa gagaggaatt ttgagtccac aatcagattt gagcgagttt 58080 gatactttcctctgattatg cttgtatgtg tgtgatatgt ttctatatat gaatatatgt 58140 atatatgtatatgtatttga ttattaatta tgagtcagtg aatactttaa accctccaat 58200 agtgtctgtcattttcatat atgtataggg cacagttatg gcttagacag tattgcttat 58260 ctgaaaatcttggaactaaa ctcgtttctc attttcgagt ttgggggaaa ttttgcgtaa 58320 actgtcctcaagtgaatcta gaaattcaac atccaaactg ctctagaatc tgaagttttt 58380 aagtgtaatgtaggcacact ttgggcatca gattttcaga tgaagagtgc ccgcctgtgg 58440 actgctgtggctctggcaac aggagtcttc acttgcttgg ccaggcaaaa gaaaaatctc 58500 gtcttcctctagcccccacc ctctcaaggc tggcccatga tttgcagcaa tcctcctgcc 58560 ccagcatcctgagtgctgag gatgtcttta ttgtccgagt aattaatgtc ctcctctgct 58620 gtgcactcaaacataaaagt tgcagtctgt ctgtttccta acattcttag ggactaatcc 58680 atgagctgtcactggggaac agctgccata aactgttgtg tcacggggga aagaaacaag 58740 tcctggtgcctgagggtttg ggcttgagat ccgagcagct tagtgaaacc acagggaatt 58800 tgtaggacagaacagaacgt gccattcatc ctggaggaag acccaagcca ttctgaaatc 58860 ctttaaaagggttgggtttt taaattcacg tgagatgcca tctagtggcc aaatagaata 58920 actgtgaaggaagccaactc atcaagatga ggttttctta gaaaaatcac atgtaatctc 58980 agtgtgattttgtgagtctt ctgtaacttt gaaagatgta tgaaaattat agtaagcatt 59040 taaaactcataagtaaaacc atttaaagaa taaaccacac aaaaagaatt tgagagcacc 59100 atgttccatcaaaacaatac aggacatagc caagagttag gtttgtaagt taggaagact 59160 tgtgtataatgctcctgcag agaccccacc ccatacacat aattacagct acctgtagct 59220 ctacttacagggacttgggc ctctggcctc tgagggcaca cgtacacggg cacaaacacg 59280 tgcgcctatatgcatacatg cacacacgtg ttatcacgca cacgtactta aatctttggg 59340 gaaaaagatgtatggtggat tcattcaaag aacttaggaa cagtaagaca gcttgagtga 59400 gagaaggcttgagggccatg gaaagaatgt ccgtgaagtc ctgggctgct gagatggctc 59460 agcaaatagaggaacttgcc gccaagcctg ggaaaccgag tttgaccccc agaaccagta 59520 tggacagagctgatttgctg acttcacaag tagtcctctg accacacaca cacacacaca 59580 cacacacacactatggcatg tacactcaca caaataaatg catttttaaa gaggaaatta 59640 taaagaaccataaagtgtga ggccaggcgt gtgtgaagac agctagaagg tgtgtcaagt 59700 ccaagagtaaagcagactgg tttaagcaaa ttcgttgtaa aataatatgt tcccttttgc 59760 acattggagatgattaataa actttgctgc tttttgtaat gggtggtatc gccttaattt 59820 cagggtgatcttttgcaagc accgtgaatg tattttggtc tcatatataa ttgattccaa 59880 gtgctttagcaggcctgtac attgcctttc tccagacatc tcagttaggg gttatccaat 59940 gcttctttagatcctgaagt tagactctaa cgtgctctgc gtgtccttat gtccgctcag 60000 ctgacagtattctggaatca gccctagcca tgtttgaatg tgtaataaca actactcaca 60060 tgttgtcctgaaacacctgg tctgttggaa atgctctgca gactccctgt agtttatatc 60120 agagtttattgcatgatggg gttcctacca cggattagtc acttctcaca ttccttctca 60180 ctgttgggggttttgtcgac ggagctggtg gcatgcagga tagcactggt ccaggctacc 60240 ttctggtgtataacttcacc ctaccaacaa cacatctcac agagaccata gatcccatag 60300 agaactgttaaccatagagc tggggtatag acttcatata actgggcagt attcggggtg 60360 tgtgtgcaagacttaagggc tgttgcttga agcaagtctg gcccagataa aaatatatta 60420 aaatgattaggtcttgaaat ctgagtagtg aataatctag atatcattaa agaaatgcac 60480 tgccagtgcttataaatata cagaaaataa aaaccaagtg tctgggggtc gttaaagcac 60540 acaaggaaagagccagtgag aatacacatt gattaatgat agcgttcttg ggaaggcaag 60600 tacgtggctaaaaagtgctc agtactttca tcttcacgca gagttcagat actccctaga 60660 cactgtgtttacatgattaa ctcagcaaac cctgggcctc tgaagggagg gagcagcgat 60720 agaattggcactagggggct gcagcctggg agggatggca tgtggctgat tctgctgtct 60780 ctagaggggacagcatgcag aagaccattt ccttcctgac cattccatat aacacaccta 60840 cacgtgtctttaacatgcca cacttcattc tgtgtgtgtt tgtatataca tgcgtatatg 60900 ggctcccttggtctgcctgc atgcttctga aggtcagata tcagcactgg gtgtcttcct 60960 cttccattctctactttgtt tttttgagac aggctcttgc aatgagctca gaacttactg 61020 ttttggctaaagtaactgat cagcaagctc ttgggatctg cctgctatgc cactgccatc 61080 cactggccccacccactggc tccacccact ggccccaccc actggccccc agcatttgcc 61140 ccacccactggcccagcact gggcttgcag acatcttctg ccatgcagca ggctgaggct 61200 ggggatctgaactcagatct ttatgcttgg atagtgagca ctttgagcca tttcctcact 61260 cgtgcttgtagactatctcc agaacaaaga gcagttgtgg acagtgaata aatcacctca 61320 gattttaccttcacgcctca ctctgaaggc caaagccaga gtctggtctg acttcagaac 61380 aaagatacccttcttccatt cattgtctgt ttcctctctt tcttcctctc ctgcctcctc 61440 ataggcttgctccgtcctct ttggagtctc acttctttat ctgcttgtta ttaaagcatg 61500 cactcggtccccagctttga aatccgacag atctttttac gtttccccct cctctcgtgc 61560 agtctcgagttgacttttaa gtttacgggt gaccccaaat gattcttctt ccagcgagtc 61620 aaggatctctaattatgaag agcaaagtgg tccctctccc ctgatggaga gcacggatgg 61680 cactcttatctgccgcagcc agggaaatga gcagggaagg ccccgggggc cataacagcg 61740 agtctataaacaagttcggt tccttaacct ccctatggcg tcggtgtgcc tgcggatggg 61800 ggcaaacggtcattagctgt aacttggggc ttcctggctt tgtgaagtct tgagaacccg 61860 catggtctgcgattgtgttc gggccgtgcg catttcccag ttttttactg tagtctgttt 61920 aaaggaatgaagagggaaac attaaatatt tattttggct gacaagataa ctgaaagctg 61980 ctagcgggaagataactggc aagtctgaaa tctgggtttg tttgtcttct agaacatgcc 62040 cattgtgtgtctttgaaagg atgatacggt cattaggcaa aaacatctta ttatgaggtt 62100 gattataatatttactcgaa gccttttgat ggcaagatcc taatcgtgaa aggaaatggg 62160 atttcttcaggaaagatgat cacatccttc ctagaaggcc agtggtaaaa gtttgtagaa 62220 aattttgcatcggctcagtc cggctggcaa gtgagcaacc ctgtgagtgg ctctgctgtc 62280 cctggttctggttctgattc ccagggagtt tgaggtgcag ttggcttctc gggaatgctg 62340 tatggctaggtacaggcatt gctccactcc tcattccccc cagctttata tcagcgggag 62400 tgtctgtgggtaccagctcc agggtccgaa atcaagtggg gacaaatcct cttgtggtaa 62460 ggatggccagctggcactag tgctctgcca gcctcacagg actgaattag ccatacttag 62520 aggttatgtcagtgggccta acctttgctc tgcaaagact tgggcctcca gcatcgcgtc 62580 tggtcagaaaacttgccttt tctctgctaa cataagagca ggtagtatac agaatggaag 62640 ggcagactgaagactgtgat gagctgctag aggtgctcct gtgggtacac acatgtgcac 62700 gtacgtgtaaacacatgtga acacgtgtgc actgagatgg ttaatcatgc ttaaggaaag 62760 tagatcgtgacccgggttcc taagtggggc tggattgtca gcatgacttc cctcactgtg 62820 gggaggcaggaggagaaggc ccacggtgta agagtccagt acatccttcc tagggcccgg 62880 gcacatggcctttaggcctc tgcaagagaa gaaggaattg gagggggttc ggtggagaag 62940 gagaggcctgcggtgactct gggagagaaa ctggttgcag atggctggtg gggagagtac 63000 atgacaccctgcaagttagc tgaaaatcag atttcttaca agcaaagact ggattctggg 63060 gagatctagaaccagggcag tttgccccag ccaagagagt agctggccta agacgagtag 63120 ctggcttgagaaggactggg tctaacagcc acatgccatt acgattggat ggaatgaata 63180 cagagacagtcactcgagca agcagagctt tttattttcc ttgtccttag cctctgtcca 63240 gagggcacagaacacagctg tgccccaagg cagtgtgctc caaggaggat ggtcataccg 63300 agctgctcccgtcagctctg gcttgtgatg agcaaagtgt gaggcatcta cgatagagga 63360 ctcgtgggactcacagcaga atgacatgcg actcacaagg tggccggtgc agtgattcta 63420 gggcaggcttttgttcactt aaatgtctcc acacagaaag cttccatgcg tcttcgtggc 63480 tctctccatattggcttttt cctgtccagt tctcaaccaa aactagacca tccagaggga 63540 acacatctacacgcggcgca ctcatgccat tttctggctt cgctgcttgt ttacctgcca 63600 ggactgatttttatgttgct tccccttagc agcctctgtc cttctccctc atggagatgg 63660 gaagagacaagctgctggag acacatctag ggtcatagat ggcagctgag cagcatccag 63720 gggtgacaccggagagaaaa cttcctccct ggctcttcct cgccttccct tctgggacac 63780 aggatttccatctaaacatt cctcagcttc aactagaatg agcttgagat gctagctagg 63840 aggcctcagcgcttgttcct gtctgaactg ggcacttgtg tggccatcaa gccaattctg 63900 tcctctctcagatctccact tacacagatg aactgggagg tggagccatg tgacccctcc 63960 tgaccccaggacctcttaga tactaggcag tagagtcctc tgagatagaa cttcaggaag 64020 ggcaggagagtgggggacgg gggcagatcc acagttgcag ctgagtctga tgggtttttt 64080 ttaaattaggtatttatttc atttacattt ccaatgttat cccaaaagtc ccccacacgc 64140 tcccacttcttggccctggc attcccctgt actgaggcat ataaagtttg cacaatcaat 64200 gggcctctctttccagtgat ggccgactag gccatcttct gattcatatg cagctagaga 64260 catgagctccggggttactg gttagttcat attgttgttc cacctatagg gttgcagatc 64320 cctttagctccttgggtact ttctctagct cctccattgg gggggccctg tgatccatcc 64380 aatagctgactgtgagcatc cacttctgtg tttgttaggc cccagcatag tctcacaaga 64440 gacagctatatctgggtcct ttcagcaaaa tcttgctagt gtatgcaatg gtgtcagcgt 64500 ttggaagcttattatgggat ggatccctgg ctatggcagt ctctagatgg cccatccttt 64560 cgtctcagctccaaactttg tctctgtaac tccttccttg ggtgttttgt tcccaattct 64620 aagaaggggcaaagtgtcca cactttggtc tttattcttc ttgagtttca tgtgtttagc 64680 aaattgtatcttatatcttg ggtattctaa gttcctgggc taatatccac ttatcagtga 64740 gtacatattgtgtgagttcg tttgtgattg ggttacctca ctcaggatga tgccctcctg 64800 atggggttttgcttgagtaa agaaatggtg tgatgaggtg tgacgatact tgatacattc 64860 accttagatcctgaagtctc tgatcgcatg ctgtgccaag gctgtttggt cttttgtcct 64920 cacctgctgcttccttttta attcagaatc ttggaactga ttggttacca ccccgaggag 64980 ctacttggacgctctgccta tgagttctac catgccctgg attcggagaa catgaccaaa 65040 agtcaccagaactgtgagtt cctagatacc ctgtgtcctc gacgtctgcc cttgagggta 65100 tgattgacaagacacggcct tggttacttc ctcccagagt taccatcttg ggtggcagat 65160 aaggtagataccttcaagat ggtcagagac tcaccgatgc caccagcccc taaatggcgt 65220 ttatgcaaattaagaaaaag actcttgagt cctcgcagat ggttaacatt ctatgcaacc 65280 ctcaggggcagtcttgaaga gcagctaaaa gtgggtgtgt aaccagcttg gggctgcact 65340 agatcggcaagcacctcggc atagaagagg gcctgggaag gttccaacga gagttggctt 65400 gcactgaatcagtttccatg gtagagagga tggggctaac cagaggaaga ggtaagagag 65460 ggggctgcctgtgcagcagc acactgtgtg catgcgggag agcctgcaca gcgaaggagt 65520 tcaggaacagagctgtccag tagtagagcc tctatcacag aaacatcctg tgccttcata 65580 ggccacacaagacctaggta gtacttgcag tgtgtctggt gcattgagaa atgtctttaa 65640 acttaagttttaaatgattt aagtagcttt tcttggcact tggctacctt attggactga 65700 gtggttctgtggtgttccct tggccacgtt tccaattcgg tctttccatt tatggatgct 65760 ccagggtaattttgaatacg taaaacccat caacatgtaa gaaactggac ttggtcctct 65820 gaggactagggattggtaga cttgatctct accctaagga gtctttgttt tggaagaaaa 65880 gggagcctggctcccagaaa ctgggtgacc tttaacctta gcatctctca tagcatcgtg 65940 gtaatacaaacagcaggtga atgataatct cataaacaga ctatatttag gaaagagatc 66000 atccaagcaggacactggta cagacagggc tgccgagggg ataggctagg agtcttcaaa 66060 taggaaaccctgagaagcca cctagcagcc atactggcaa tgtaggaaac agaagtgtag 66120 ctttctgttccaagtctgat tctgcccgac actcctctgt ttacagtcgg gagagctcga 66180 gctcagcttcactaactgtg cagcaactcc acaagctgtg cacagcatgg ggggggggga 66240 gggagagagagagagagaga gagagggagg gagggaggga gggagggagg gagggaatat 66300 gagaattcaaactcacccct ctgtttcctc cctgggagct ttgggtttca tctgatctaa 66360 taatacagcaggggccatta tggcatctca gacaagtctc ttactgtctc tctggcataa 66420 aaacaaaaatatccaaatag actaccccag gggtggcagg acactccctg atgctctcag 66480 ggagatgacccagccagaga ctccaaggta tggtgtcagc cttctcttgg agggccccca 66540 atgcttttgtgctccctagg ggttccccac ccccaaggcc gattggtaga gacacacaac 66600 tgtattctgctcagctctgg cctagctcac cagctagtgt ctctgggtct ggcttccttt 66660 tttttcttggccttctaagc caatggggtc tgtcaacaca gccgggaaaa tatcttcatt 66720 ttaagtacctgggaattctt ggagggtagt gttatacttt gtctcctggg gagggccgtt 66780 agagagctcagtggccatgt ggaaagaaga cggctccctg tttagagctc cctcatctat 66840 aaagagggatagcaaggaag ctggtcctac cggtttgcag ctgactcctg ggggaaatac 66900 gttggaaaaacgttttggtt ggatattgtt tttacttaat ccatatattt aaccccccag 66960 cacacctaaggataactcac agtccaaata gaaagtacag actcttgatt ttctgggttt 67020 tgtttatttttttcccccag tggagttagg aaaagctgcc ctgagcacag aggcctgtat 67080 ttaaccaagaggcctgcatc aagcaaagtc tttccgccct atccccctac aagtacagtt 67140 ccattctagatcccctgcca gtgtgtgggg tctctgattc accatgtgtg tcacagcttg 67200 tgtgtgtttcccatccaaac gcctggagaa gctccgtcct cagccagatt tagataattg 67260 aaagtcatcatttccaaatg ggctaataat ctatacactg tcactctcag aggggaagag 67320 ttgggctggtttctctttgt tgtgctctta ggaaaacaag attaaaggcc ataaaaagcc 67380 tccctctctctcctggagtg gcaggccctt caccaaggct ggcattaatg agaggaatgg 67440 gagtggtgtgacttagaggt gactgataca gcaggcccgc gcctgcactt aacagaagtg 67500 cagctcaaacctccgctcag cgaacttctt cgtgaaaaag actaagaaca ctgaatgtgg 67560 caagcaataagaactatgta aatgagctaa ttccaggctt gatgtcatgc aagaattgcg 67620 tcactgcagccaggtggcct ccatgtcacc cagaaagcga cagcagaaaa gataaaggtt 67680 ccttctctggccaaggagtc catgatggtt aacaacatgg ccttagcccg tttttttttt 67740 tttttttcttctcacttctt tctggctcaa cataatgtag tcagagagga tccatggccc 67800 ctggaatcaaaagaccgatg atctaaacac tatgactgtt attctgtgca aagtgaagca 67860 aacacagccttcctaggctg cgtttccttg tgttagaggg tgctgtctct cctccctcct 67920 gcacaggagtccatttaaaa gattagatga agataagaca atcaaagtgg caaaacaatc 67980 tgactacttctgataataaa aacatggtct tcagcagata atctagaagt ttcttacctt 68040 gttacacccccatgaaaaag aaagccacgt tcttttggat gtggatttgg atacagatgt 68100 ctattgcacttttattcaca acgggaaact caaacatcca ggtcaaagga ataaactggc 68160 catggtacacccagggagac acttctctcc cacgaatcac agaaacaccc acaacctcag 68220 gaacacgatgctcaaggata atgaagccaa actcaagaaa gtctactccc atagacgtct 68280 agaaaggacaggagacggtg ccggggaaag atcggtgatc gtctggaatc agaagtcaga 68340 actaggaggaggagttaggc acgtgggcca tggggcaggt tcagatgaag tttttggtcc 68400 ctctgctgtggtgatggcta cttaggtgtt tacgtttgtc aaaactcact gtactatatg 68460 cgtgaaatagttacctctcc atgggattaa ttaaaattaa ttaattaaaa ttaaaattaa 68520 ttaaaatacaataaagcttg ttgtagagag gaaaatgtgt ctaatctaga ctctcaagcc 68580 tgagactgtgcaagctggca atcatgttta gccaaaggac tctttcaagg gctgcagtcg 68640 ctgtcgatgggaagcaggag tttcacacac tggtaaccca caccaagcaa ggcagccatg 68700 acactgaactccacaggtgc actatgtaat gactgtcaca gggacttgaa tttacctctc 68760 aaggcagcggtggctctgct gagcaatgcg cacatgtaat tctccatcat tcctgctcct 68820 cgctgctgtgtgttaagata tccaagaggg taagatgtgc caaggcagaa ggactgtgac 68880 tggtgtgactatgaacaagg ccatccctca tcagaccatg caggactgat gaggtggccc 68940 acgagctgactggtagatcc aaggagactg cctacaggga acatagctgt tctctcctag 69000 aaccccaggacaattgccgt aaacaattcc tagtaacata actggcacag tacctaagca 69060 ctaggcattgatagcttgtg gaagaggaaa ttcctaaaca gtgtttgtgt agccgatgag 69120 gcaggcaagtccatttgatc tgcaaacagc agtagccttc tggacagtta gaaatgacag 69180 tgatggacatgagaggaaac cacaccagga ttgttactta cccaaagtcc ccaagccagc 69240 aagacttcctgagcctccca tcattcctct tcagttctct gtgacctggg cacttctggg 69300 ctggagctctgaacaatcat ttggaggggg gggggggtga aagtcagtgg acagtttaag 69360 aagctaacagacatttctaa aatcagcaga catgactgaa aaggagccct tgagagctct 69420 ttcacaaaaataaaaatgca tttggaaatg gggaagaaga caaactagtg tgtgggacac 69480 cccttggtgtttttagactg tgaaattgac tacttagaat ggcctaggag aggaaagaaa 69540 ctctaagctggcagtctcac ggggcagttg gcaagtttaa agaccttcca gtggctgagg 69600 ccctgaattaacagtacagg cctttcccta ctccagataa atagagcagt agtttctctc 69660 aaattgtatctggctcctaa ggtagcagaa agttcaaggg acacaaagtt taaaaaacaa 69720 acttcagctgggtcttgtga cacagctctt gagttgagtc ccagttattc agggggctaa 69780 ggcagaagaatggtttaagg cccgcctaga cttcagaggg agttcaagga caacctgact 69840 aatgcatcaaaataacagaa aggactgggg ctggagctcc gtggacgagc acttgcccag 69900 ccctgttctgtcctccacag tcagggaatg ggacggggct gcagccctgc ggtccacctt 69960 ccagatgtggtctgctacct tcaggaggtg aagtcagtag ggtgctgctc cctggttgac 70020 tgggagtcactcctgcactg ggcaggcaaa catctgcttt ccttcagaac tcctattccc 70080 attttttgaaacgaagagtt ggctgaaatg atctccccct cccccaaccc tctcactgtt 70140 cctggccttggcatccatct ggcagctctg tccatccccc tccccgggca ggtgacactt 70200 cctgtggggttagagtgtgt gctgcacacc actcctgcac ctctcttacc tctggctcct 70260 ttcttcactagtgtgcacca aggggcaggt ggtatctggc cagtaccgga tgctagccaa 70320 acacggaggatatgtgtggc tggagaccca ggggacggtc atctacaacc cccgcaacct 70380 gcagcctcagtgtatcatgt gtgtcaacta tgtgctgagg tgagtcgagg agggagcagc 70440 cagcctctcgggaccctggg cagtacctcc acatgctggc tgtggtgtgg atccttctaa 70500 ctgggagggctctatattga ggtcttagga acagagctct agcgtgtttc tttttcattt 70560 gtcagctatggtatcatgtc tctgacttgt gctgaggggg gagaagatgg catagagtaa 70620 tgcttgatgttgtgtgaatg ggagacagtg ctctcctggg taagctgttt tgggaagatg 70680 tggatgttataccctctccc ctccacccct ggaaagacta tgagcggctg ccattgataa 70740 tatgaaggaaactaacatag gaatccagga cttttgtgtg cctacagctt gagcacaggg 70800 agcgagcgaacagcccacag gttgaggcta acctcaccgt ctctgcttct gtggctcaca 70860 gaattggattgtttgtgttt ctgcctaaca cttctctgag aggtttatct accaggcttc 70920 tgacatgtctggggcgggag agccaaaggc ttttgctaaa agatgcagat actctgagcc 70980 gctctctcttcccatcagtg agatcgagaa gaacgacgtg gtgttctcca tggaccagac 71040 cgaatccctgttcaagccac acctgatggc catgaacagc atctttgaca gcagtgacga 71100 tgtggctgtaactgagaaga gcaactacct gttcaccaaa ctgaaggagg agcccgagga 71160 actggcccagttggccccca ccccaggaga tgccattatt tctctcgatt tcggtgcgta 71220 cttcctagccctggttgaac ccacagaacc ctcatggact ggcggacagt tcttgttatg 71280 acaagcctccctggccacag cttccctaaa ccacagatgc actcgggcct tgctgatcac 71340 tgtgcgtggtcaggttctgc taggtagaga agaagcacag actcatggcc actgagttat 71400 aagtcctcatgaagggtaaa gaggtaggca aggagagggc tccctctcga ggggcccatc 71460 ctctacctggcttgagagtc tgagttgagg cgtgtgtctc agggagtgtc ctctaactta 71520 agcgaaaggctgagtcagaa cagcgccaga accagtagcc gagaggagac tgagggcgag 71580 gatgaagccctcggtggctg cctgcgttag aatgtctgcc catcttcctg gaagagaagt 71640 ccttttggtcctgagctctc ctctgacacc cggaactgtc cagggagagg cctcttccgg 71700 tctgcaccctctccagccca gctctgactg ctcccctttg caatcaaatc cccctttgat 71760 aatgtgcatctgagaggcca caggaaaatg gacacctcag agaaccagaa agggcaatga 71820 gccctcttgcacgagataac cttgtaaccc cccagctcca tgttggtact gaggcaaatg 71880 gcccaattctcctctgataa cttcctcagc tctgttctgg aaggctctgc aggaaacaac 71940 tgcttctatctagtaagctc ggtctctgaa tgccaaatgc tgctggagat tgctctcttt 72000 ataggaagtccccagattga atcatagttc tggtcctatc taggctccac agtactgagg 72060 gtgtcaacttcaggccctct tttaggacct tttgtgaact ttctgggctt atccaccctg 72120 gggtgagcccagatctcacg aatgctcctc aagcaagcct tgtctttaca ggaagccaga 72180 acttcgatgaaccctcagcc tatggcaagg ccatccttcc cccgggccag ccatgggtct 72240 cggggctgaggagccacagt gcccagagcg agtccgggag cctgccagcc ttcactgtgc 72300 cccaggcagacaccccaggg aacactacac ccagtgcttc aagcagcagt agctgctcca 72360 cggtgagcccccaccctcca ggagagcaca cagggctcat gggcccctca agctctgctg 72420 ccagatgactggacagaccc cctgagaagt actgcctccc ttgggtgtta cagtgcccct 72480 aaggatggctcagatactcc gagagacact agagctgact accggctctt agcatctgtc 72540 ttccacccttatccacgtcg tgactcttga aatcacagca agaacctagg caggtctctg 72600 caaaccaaaggcttcaacca cagagccctg ctggccacag cctctatcca gtttgcacat 72660 caagacacacaggacaagta gctcacaacc cgtggtacac aaatccctct atttgacggg 72720 agaaatcgaacaagcctttc acagggtcac ctagatcatc agaaaacaca gaaattcact 72780 ttataattcataactgtagc caaattactg ttttgaggtt gggggtcagc acagcatgag 72840 gaactggagggctgcagcat caggaaagtt aaggaccact gatctaggaa gtgatccaag 72900 cctctccttagagggagacg aaggttcaga gaggttgatg agctgagctg cttcacacag 72960 cttgtaaatagtagtcccct tcacagatgc ttggtagaac ggggcagagt gcagaaccat 73020 gcttgtctgttctcaatgga ctcctggcga gacaggcccc gtgttcttac ttggtctcgt 73080 ccttcctggtctccagccca gcagccctga ggactactat tcatccttgg agaatccctt 73140 gaagatcgaagtgattgaga agcttttcgc catggacacg gagccgaggg acccgggcag 73200 tacccaggtgggccccgcgc gtgggtgaca gagggctcct ttgcagagac ccccggtgtg 73260 ctgcgcaaagcttcggggtc aggaagcttg tgagtggcgc gctcccttcc tactgtgtcc 73320 ctgctcagccccacactcct gtctaactgt tgaccccatc tctgtcttgc acaaatggtg 73380 taagtggtggtggtggaggt gggcgagggt ggtgatgaga atctccaggg gcgatcctca 73440 cacctccctttgattgtctc agcctgagtt ctccacccca gcttccacat tgttctttag 73500 tgtgtggccttatcctatcc tgcagaacct cttgttgttt ctgtgcatgt ctttttcctc 73560 tgtgtgtgtgtgtgtgtacg cgcgcgcgtg tgcctgtgac acttcttagc acaagcaggc 73620 acacttccttcagtaccagt cttgggtaca aagatctatc tgtagtgagg ggtcattggc 73680 tgaccgaccgcggctgaagc ttgtgtcttt gcttatgagt attctgttgt gaccatgggt 73740 gtccctgagtatctactggt gagtgagaac agcagtccct gtgggggaga tgaagattag 73800 ctggtacaaataggataaag gagcagtgca acagaggcca gaagttggcc tggctaaaag 73860 agaaagagaaggctgcaggt caaaagtgga gaaagctcac tgaacctgta caggatgaag 73920 ggacagatgcaggttatatc ccaacagcaa tccttgtacc ctcaatggca aatctgagca 73980 gttccagcagaggtatttga atggaggact gacagtttca aaaggcctag gcagggcagg 74040 gctgggccagccactatgta ccctttgctc tgtgtctcct cagacggact tcagtgaact 74100 ggatttggagaccttggcac cctacatccc tatggacggc gaggacttcc agctgagccc 74160 catctgcccagaggagccgc tcatgccaga gagcccccag cccacccccc agcactgctt 74220 cagtaccatgaccagcatct tccagccgct caccccgggg gccacccacg gccccttctt 74280 cctcgataagtacccgcagc agttggaaag caggaagaca gagtctgagc actggcccat 74340 gtcttccatcttctttgatg ctgggagcaa agggtccctg tctccatgct gtggccaggc 74400 cagcacccctctctcttcta tgggaggcag atccaacacg cagtggcccc cggatccacc 74460 attacatttcggccctacta agtggcctgt gggtgatcag agtgctgaat ccctgggagc 74520 cctgccggtggggtcatcgc agttggaacc tccgagcgcc ccgcctcatg tctccatgtt 74580 caagatgaggttagtgacag atgtctggct ggaggggaca tactgggcag ggcgagtaca 74640 tgtacacacctgcttatgaa ggctctagag ggtcacatca gcttcccgct gaccatatcc 74700 cctcactgtatgtgtaggca gctggcactt ttcctagtcc tcctctgata tgtgagagcc 74760 agaccccagcagatagaaga aggggctttt ttaagtgatg tctgtctacc tttggacaca 74820 gataggtattatggtatttc ctccatagtc ctgtactttc catctgcctg gagcctctgt 74880 ggggtgcacacaggcaccag cattttcctg tcttcccagt aggctgcagt cccataagct 74940 gatgtgtcttgtctatagta cgtacccttt atagagcacc ccagatggtc ctctggaagg 75000 aaggcattcagcagaaatcc ttctagagac tgttgtcatc tagctgggca ggacccctga 75060 gggcaggactgaggtggaca ggtgtgcgga gagaggcgca ggtgacagaa ctcttgagca 75120 tcttacagaaggaagagctg agctgccgcc gctcaagccc tttagcattc tgccctccat 75180 agatcgcctgagtcataagt gttagattct gatgaagaca ccaggcagcg ggaggctgta 75240 gatgcctcacacccaccact gtcttctctt ggcattggca ggtctgcaaa ggacttcggg 75300 gcccgaggtccatacatgat gagcccagcc atgatcgccc tgtccaacaa gctgaagcta 75360 aagcggcagctggagtatga ggagcaagcc ttccaagaca caagcggggt aagccatgtc 75420 tgtgaacgaacagcctactg gacaggaagg agatgaggct agggtagaga tgcagacagc 75480 tagatctggtagtgagcacc tgcccctgct gggtctgtgt gctacccacc ctactccacc 75540 ctggcccacttcctccaact acacatcacc tctgcagggg gaccctccag gcaccagcag 75600 ttcacacttgatgtggaaac gtatgaagag cctcatgggc gggacctgtc ctttgatgcc 75660 tgacaagaccatcagtgcga acatggcccc cggtaagcag gcctggccca ggggtctggt 75720 ggagggttgaaggctcagag cacattccct gagccttgtt agaatgggtt atatccatgc 75780 catgagcaggatcccggttc agaggtctct acatgacttc tgaaaaagaa agcaggctga 75840 gagcgctattgtctgcccta atgacagcac cacagttcac tctctggctt ggatcctcca 75900 gatgaattcacccaaaaatc tatgagaggc ctgggccagc cactgagaca cctgccacct 75960 ccccagccaccatctaccag gagctcaggg gagaacgcca agactgggtt cccgccacag 76020 tgctatgcctcccagttcca ggactacggt cctccaggag ctcaaaaggt gtcaggtgag 76080 tgctttggaactcccaccat agccagggtc tgatgcaaac aggtggcctg gctggccaga 76140 gagctgagaggcagccactt gcaggaagca actctgcgtt tggacaagac tgccttcaga 76200 gcccctgctcaggtctgatg cgactgctta cagtccatag ggctcttcat cagccaagct 76260 gcatgagatggtttaggcag gaccaagtct cctctttatt ccagagcctg catggtacca 76320 ggcccagaaaggatttgaca atgacttgtt caaagtacaa ggaaggccca ggagggtgca 76380 ggctcctcattgatgagctg agattgtgtg agaaagtaag aaaggcctct ttggctgttg 76440 tccctgagaaagggcagagc cacggttact tctggccaag caggctcccc agaaggtggg 76500 tctcaggctggggaagcttt ggtagaagga agcggctggt atggcaaggg ccccagggct 76560 gtggtggacttctgagcaag acaagctcgc tgccctggaa aggcaggctg gaagattgtg 76620 cacggctctggaagggatcg tccagtaatc agtcagatca catccgggct cttggccccg 76680 caagaccaagagtctacatc tttcctcagc tgctcattgc ctgaagtgta tacacatacc 76740 gtaacaagctttctctgaaa ggttgcacag gggactcact gatgggaggg tagcctggtg 76800 atccagtaagttcagctatt gccactgaaa cacacaagtg ttcccataga cgcctttgct 76860 cggcacaaacaatctgtggt ttgagaaaga aaattccaga gccttgtcta cagtagcaga 76920 gttggggaaaatttatagtt gtgttcttag aattccatct gaagaaccca aaagccctgg 76980 gctgatatttcagttctgtt gtttaccagc aggtgggctt gcaagccaga agtcttaaaa 77040 gagaaagtttaaagtttaaa gagaggacca cacatcactc acccccggta tacacatctg 77100 ggcagctgtagtcagcaaat gacaggtttc cccagctgtt gaatgcgaag caaaactaat 77160 ttcaacatagatcgtttctg cccatagacc gaaaggaggt gaatttaaga agggcagggc 77220 atctgttggtaggagtgcag gcaacgggaa gtgtggaccc cagaagggag agcacactgg 77280 gacccagctctgtctctaaa ctgagagagc tctgctctcg ctccagccat ctttggatct 77340 caccactgctgtggcatgtc ctcttctggc tccgtcctct gaacagggga tggagcccca 77400 gggccagagtgctgctgcat cggaagttgc tggaagctgg agacacagtt gtttgcatgc 77460 atgccctttcctctggtcat ctacagccag cggagcaggt gtgtgccagt cctctaaaac 77520 accctcctctccctcctctc aggcgtggcc agtcgactgc tggggccatc gttcgagcct 77580 tacctgttgccggaactgac cagatatgac tgtgaggtga acgtgcccgt gcctggaagc 77640 tccacactcctgcaggggag agaccttctc agagctctgg accaggccac ctgagccagg 77700 gcctctggccgggcatgccc ctgcctgccc cgccgtcttg acctgccagc ttcacttcca 77760 tctgtgttgctattaggtat ctctaacacc agcacacttc ttacgagatg tactcaacct 77820 ggcctactggccaggtcacc aagcagtggc ctttatctga catgctcact ttattatcca 77880 tgttttaaaaatacatagtt gttgtacctg ctatgtttta ccgttgatga aagtgttctg 77940 aaattttataagatttcccc ctccctccct cccttgaatt acttctaatt tatattcccc 78000 aaaggtttttctctctctca ttcatatcca tactaacaag catggtggct ggtgcctctc 78060 cctaggaaagctttggcgtc attcaactca agtgttcttg ttcttgttgc caaagagaaa 78120 aggattttcctccactgtgg attctccctc tcccccaccc ccacatacac acacacacac 78180 acacacccctacacacatat acacacatgc acgtatgcgt gcacacacac acacacacac 78240 acacacacacacacacaccc ctacacacac acacacacac acacatatac acacacacac 78300 acacacacacacacacccct acacacatat acacacatgc acgtatgcgt gcacacacac 78360 acacacacacacacacaccc ctacacacat atacacacat gcacgtatgc gtgcacacac 78420 acacacacacacacacatct aatcaccata ttgtaaaatt ttgtgttttt aaagccaact 78480 ctttgctccggttttttcat acgacttagt atggggcaaa aaagcaatgt gaagaatcaa 78540 ctctagggttacctgtgaag ccacgcggtg gtgttcgaag ctgtctggta atgcccccat 78600 ctctccccgggtccagtgga tttttttaac tattattcaa aagcaaaact gagttttgtt 78660 ttgtttggttttttaagaag aatttatatc cgggt 78695 <210> SEQ ID NO 258 <211> LENGTH: 379<212> TYPE: DNA <213> ORGANISM: mus musculus <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: (345)..(345) <223> OTHERINFORMATION: n is a, c, g, or t <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (367)..(367) <223> OTHER INFORMATION: n isa, c, g, or t <300> PUBLICATION INFORMATION: <301> AUTHORS: Aizawa etal. <302> TITLE: Computational Analysis Of Full-Length Mouse cDNAsCompared With Human Genome Sequences <303> JOURNAL: Mamm. Genome. <304>VOLUME: 12 <306> PAGES: 673-677 <307> DATE: 2001 <308> DATABASEACCESSION NUMBER: BY229956.1 <309> DATABASE ENTRY DATE: 2002-12-10 <313>RELEVANT RESIDUES: (1)..(379) <400> SEQUENCE: 258 gattcgagag cggccggtgtacagctccgg agtccgcagc gctccgctcc agctctcctg 60 aggcggccgt acaatcctcggcagtgtcct gagactgtat ggtcatctca gcggccgcac 120 tcgcttgccc ccggatttttttccaacttg ctctcttcga gccatttttt tttctttttt 180 tctttttctt tttttctttttctttttggg tgggttggtt tggatttgtc agatcccaga 240 aaagtgactc ctgttcggggctaaacggaa ctccaggtcc cttgtgctgc tctctctctc 300 tttgggcgtc ttacaacctcctcccactcc tttccccggc cccgnctcct cctgcaggtt 360 cctcccngtc atcccccta 379<210> SEQ ID NO 259 <211> LENGTH: 2730 <212> TYPE: DNA <213> ORGANISM:mus musculus <300> PUBLICATION INFORMATION: <301> AUTHORS: Carninci etal. <302> TITLE: High-Efficiency Full-Length cDNA Cloning <303> JOURNAL:Meth. Enzymol. <304> VOLUME: 303 <306> PAGES: 19-44 <307> DATE: 1999<308> DATABASE ACCESSION NUMBER: AK087208.01 <309> DATABASE ENTRY DATE:2004-04-03 <313> RELEVANT RESIDUES: (1)..(2730) <400> SEQUENCE: 259gaccatcgta aaaatgtttg ctacttaact gcactccctg tgtagcacac aggaagtgct 60gtgtgggacc tgcacagtgt tttgaggaca tgattgccct ctgttgcgga taggttgtct 120tttcatggac agattgttgc taatgtttct ttatagtgga atgtgcccag gactaaaagt 180ttcacataaa taaatggtca cagtatgtcc tcacagttac tggttactga tgcgacactt 240aggcagcttc atggtagaat ctgacgagtt agcaggcaga tactctgact tttaaactta 300cccgtgttag tacgtgatat ggactttgta cgaagaccgt gtttctttag gatctctgga 360aagaggcagg tttgggtgtc agtttgtcct ttccttccca ttctgcaaca aagaagagtc 420agtctggcac ctcaggctgg caaggatggc acccactgca gctaccaccc ttggaggtct 480ttgcttctgg attgcaaatg gaggcgtgtt gtccgcctca tgttctcttg gcctttactg 540atgtctccag actctaacct gtcgtctctc agatcagaaa cagggtctta ggtaagccag 600ggcctggtct gaccgtagct tcttcgccct tctctttcca ttggtgccct ttgaccctgt 660cctcaaactt tgttcattag tttaattaaa tctttgctaa cgctacccac gtgaagccca 720gttctggctc ctgcaagaat acagaagaaa gcaatttgag aagacaccaa tgcgcaaaag 780cagagtcaat accaaaaggt ggcttgctca tagctcccct gggctgagcc agatgggttc 840agtgggagaa ttgactcact gtgggggtga gtgggtcact accgagagtg tgaatggatg 900acgtccacat tccaggacta acccctcgtt tcttcatgta ggagcagctc agagctgagg 960aaggagaaat cccgtgatgc cgcgaggtgc cggcgcagca aggagacgga ggtcttctat 1020gagttggctc atgagttgcc cctgcctcac agtgtgagct cccacctgga caaagcctcc 1080atcatgcgcc tggccatcag cttccttcgg acacataagc tcctgtcctc aggtaaggct 1140tgacaggtcc tgcccccaag ctggcatcta cctaggcctc gctccaagac acatctacca 1200atatccactc acagaagctg gcacatggcc tttagtgtta catttattta gttgcgtgtg 1260agggtatgca tgtgggtcag aggacagcct ttgggagtcc attctgttct cttcttccat 1320catctgggat ctgggacttg aacttgggtc ctcaggctta gcagcaaatg cctctagcca 1380ctggaccttc ttgctggccc tgttccttca ttttagcatc tcccctctgg caatgatctt 1440ctcatgagtt cacccaggga agagaccaag gacagactca agtgagagtg tgaggtgctc 1500ccagagagtg tgaggtgctc ccagagagtg tgaggtgctc caaggggttg gagagccgag 1560agcagcttct cctggaagcc catccagtac ctctggacct ctggcgagag tcccgctcca 1620cactgtgttg actctgcagg aagcctttta tccttgtctt ccagctacat ctctaggaca 1680tcagaaatgg tgatgtccct tgtgatctat ctctcagaac cttggtttcc ttgcctacaa 1740actggaatta gccaggcata ctgcctggga ggataagggg taggaaatgg ggggggggga 1800ttattagggc actataggaa tgagtggaga ccgcgggtca gctgtattcg ttcttgctgg 1860ggctagcccc ccccatagag gacagcctcg ggcacctctc cctgggtcag ccgatgcgtt 1920cttctttccc gcatatctct tcacccacca accgttcata acgaatgctt tctttccttt 1980gtcagagtta catccctcaa aaatcatttc ctgttaggcc tcaccaggaa gaggcagcct 2040gggggttcca ctttcacatc ctatgtgcag tcttgtcaga cttatcagtt ctgtaaggaa 2100actgggcagc atatagctgc caggctggca ctacagcagg gcagtgtccg aggcatgagc 2160aagggaggca ggcaggcaag ggggaaagag atcccctggc tcattttgag ttttcctgag 2220tgagtgtgtc actctggaga tgactcctta catggctatt ctgggaaaga gccccctgca 2280cagaggggtc cagaatgagg cggggaagcc agactagcct gtgctattct gggcccctgt 2340gcacaggaag gatatatggg aaagaccttc ggaggttaga atggctgctc atcccatcgt 2400cctcctctaa cccccaggct ggaggctaag cctgggctgc aaggctgagg tgaccgtgct 2460gttacagaaa tgagcagaga gtggagaaag caagggcgga gccgctgcac acacagcagg 2520gcaacagcaa ttactcagat ttagacggtg aaaatggttg agggaagctc aggctaagga 2580cttgtaaagc ctggactgct aaataaaaag gcagactcgg aggtgtctca cccatgcccc 2640atgcatgcct tcattttaca gaggattgtc ctcttggaga aatgaggacg acagttcggt 2700gatttgtagg attttgcaaa gcctgtcagg 2730 <210> SEQ ID NO 260 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Oligonucleotide primer <400> SEQUENCE: 260ggttccttaa ccccgtaggg 20 <210> SEQ ID NO 261 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Oligonucleotide primer <400> SEQUENCE: 261 acctgggttccttaaccccg 20 <210> SEQ ID NO 262 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 262 ggagcacctg ggttccttaa 20<210> SEQ ID NO 263 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 263 ttgtcagctg tcattgtcgc 20<210> SEQ ID NO 264 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 264 tctccttgtc agctgtcatt 20<210> SEQ ID NO 265 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 265 gaagacctcc gtctccttgc 20<210> SEQ ID NO 266 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 266 caggtgggag ctcacactgt 20<210> SEQ ID NO 267 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 267 aagctgatgg ccaggcgcat 20<210> SEQ ID NO 268 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 268 ttcaggtaca agttatccat 20<210> SEQ ID NO 269 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 269 aaggctttca ggtacaagtt 20<210> SEQ ID NO 270 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 270 aatgaaaccc tccaaggctt 20<210> SEQ ID NO 271 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 271 atgaacttgc tgatgttttc 20<210> SEQ ID NO 272 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 272 gtcccatgaa cttgctgatg 20<210> SEQ ID NO 273 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 273 acctgggtaa gtcccatgaa 20<210> SEQ ID NO 274 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 274 tgttagttct acctgggtaa 20<210> SEQ ID NO 275 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 275 gtcaaagatg ctgtgtcctg 20<210> SEQ ID NO 276 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 276 ggatgagtga agtcaaagat 20<210> SEQ ID NO 277 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 277 atgaagaagt cacgctcggt 20<210> SEQ ID NO 278 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 278 ttcatcctca tgaagaagtc 20<210> SEQ ID NO 279 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 279 tgcacttcat cctcatgaag 20<210> SEQ ID NO 280 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 280 tgacagtccg gcctctgttg 20<210> SEQ ID NO 281 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 281 actctcactt gcccggtgca 20<210> SEQ ID NO 282 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 282 gttgttgtag actctcactt 20<210> SEQ ID NO 283 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 283 attggctcac acatgatgat 20<210> SEQ ID NO 284 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 284 tgggtgctgg attggctcac 20<210> SEQ ID NO 285 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 285 atgctgtggc ggctcaggaa 20<210> SEQ ID NO 286 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 286 gggtggtaac caatcagttc 20<210> SEQ ID NO 287 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 287 gtgcacaagt tctggtgact 20<210> SEQ ID NO 288 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 288 ccttggtgca caagttctgg 20<210> SEQ ID NO 289 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 289 gtcccctggg tctccagcca 20<210> SEQ ID NO 290 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 290 tgaccgtccc ctgggtctcc 20<210> SEQ ID NO 291 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 291 gtagatgacc gtcccctggg 20<210> SEQ ID NO 292 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 292 gggttgtaga tgaccgtccc 20<210> SEQ ID NO 293 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 293 catagttgac acacatgata 20<210> SEQ ID NO 294 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 294 tccatggaga acaccacgtc 20<210> SEQ ID NO 295 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 295 tctggtccat ggagaacacc 20<210> SEQ ID NO 296 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 296 aaagatgctg ttcatggcca 20<210> SEQ ID NO 297 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 297 tggtgaacag gtagttgctc 20<210> SEQ ID NO 298 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 298 agctcctcgg gctcctcctt 20<210> SEQ ID NO 299 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 299 ggccttgcca taggctgagg 20<210> SEQ ID NO 300 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 300 aggatggcct tgccataggc 20<210> SEQ ID NO 301 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 301 ctgctgggcg tggagcagct 20<210> SEQ ID NO 302 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 302 tgaagtccgt ctgggtactg 20<210> SEQ ID NO 303 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 303 tccaactgct gcgggtactt 20<210> SEQ ID NO 304 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 304 ttgctcccag catcaaagaa 20<210> SEQ ID NO 305 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 305 cagggaccct ttgctcccag 20<210> SEQ ID NO 306 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 306 gtgctggcct ggccacagca 20<210> SEQ ID NO 307 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 307 cttgaacatg gagacatgag 20<210> SEQ ID NO 308 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 308 cagacctcat cttgaacatg 20<210> SEQ ID NO 309 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 309 ctttgcagac ctcatcttga 20<210> SEQ ID NO 310 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 310 ttcagcttgt tggacagggc 20<210> SEQ ID NO 311 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 311 gtgaactgct ggtgcctgga 20<210> SEQ ID NO 312 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 312 cacatcaagt gtgaactgct 20<210> SEQ ID NO 313 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 313 ccgcccatga ggctcttcat 20<210> SEQ ID NO 314 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 314 aggacaggtc ccgcccatga 20<210> SEQ ID NO 315 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 315 caggcatcaa aggacaggtc 20<210> SEQ ID NO 316 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 316 gatttttggg tgaattcatc 20<210> SEQ ID NO 317 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 317 ctggccacgc ctgacacctt 20<210> SEQ ID NO 318 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 318 gatggcccca gcagtcgact 20<210> SEQ ID NO 319 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 319 cgaacgatgg ccccagcagt 20<210> SEQ ID NO 320 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 320 aggtaaggct cgaacgatgg 20<210> SEQ ID NO 321 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 321 cagtcatatc tggtcagttc 20<210> SEQ ID NO 322 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 322 cctcacagtc atatctggtc 20<210> SEQ ID NO 323 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 323 gttcacctca cagtcatatc 20<210> SEQ ID NO 324 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 324 ggcacgttca cctcacagtc 20<210> SEQ ID NO 325 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 325 gcacgggcac gttcacctca 20<210> SEQ ID NO 326 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 326 tctctcccct gcaggagtgt 20<210> SEQ ID NO 327 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 327 tctgagaagg tctctcccct 20<210> SEQ ID NO 328 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 328 ggtccagagc tctgagaagg 20<210> SEQ ID NO 329 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 329 gctcaggtgg cctggtccag 20<210> SEQ ID NO 330 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 330 ggccctggct caggtggcct 20<210> SEQ ID NO 331 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 331 agaacaagaa cacttgagtt 20<210> SEQ ID NO 332 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 332 aacagttgag acatgacagt 20<210> SEQ ID NO 333 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 333 tgtcactaac ctcatcttga 20<210> SEQ ID NO 334 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 334 acaggagtca cttttctggg 20<210> SEQ ID NO 335 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 335 catacagtct caggacactg 20<210> SEQ ID NO 336 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Oligonucleotide primer <400> SEQUENCE: 336 aatctgtcca tgaaaagaca 20<210> SEQ ID NO 337 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 337 cagcgacaat gacagctgac 20 <210> SEQ IDNO 338 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 338 caggccacct gagccaggcc 20 <210> SEQ ID NO 339 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:339 tagactccga gaacatgacc 20 <210> SEQ ID NO 340 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 340 agcagcagcagctgctccac 20 <210> SEQ ID NO 341 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 341 ccactgagcg caaatgtacc 20<210> SEQ ID NO 342 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 342 agaagagtaa cttcctattc 20 <210> SEQ IDNO 343 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 343 atggacgggg aagacttcca 20 <210> SEQ ID NO 344 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:344 ttaccaccct gaggagctgc 20 <210> SEQ ID NO 345 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 345 agcctatgaattctaccatg 20 <210> SEQ ID NO 346 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 346 cgacctgaag attgaagtga 20<210> SEQ ID NO 347 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 347 gtgcccgtgc tgggaagctc 20 <210> SEQ IDNO 348 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 348 tgggagcctg cctgccttca 20 <210> SEQ ID NO 349 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:349 gctgtggcca ggccagcacc 20 <210> SEQ ID NO 350 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 350 tgcgaccatgaggagattcg 20 <210> SEQ ID NO 351 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 351 cctgatggcc atgaacagca 20<210> SEQ ID NO 352 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 352 ggccaaggac caatgcagta 20 <210> SEQ IDNO 353 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 353 acccagagcg aggctgggag 20 <210> SEQ ID NO 354 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:354 ctgggaagct ccacgctcct 20 <210> SEQ ID NO 355 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 355 aagcaaagacatgtccacag 20 <210> SEQ ID NO 356 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 356 gagctggact tggagacact 20<210> SEQ ID NO 357 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 357 aactgccctc ctcacaatag 20 <210> SEQ IDNO 358 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 358 ggtggcagca cctcacattt 20 <210> SEQ ID NO 359 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:359 agcagctgct ccacgcccaa 20 <210> SEQ ID NO 360 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 360 agagttcttgggagcagcgc 20 <210> SEQ ID NO 361 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 361 atttgagtcc tacctgctgc 20<210> SEQ ID NO 362 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 362 gacccacctg gtggcagcac 20 <210> SEQ IDNO 363 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 363 agactccgag aacatgacca 20 <210> SEQ ID NO 364 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:364 ttctccatgg accagactga 20 <210> SEQ ID NO 365 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 365 ccatgaggagattcgtgaga 20 <210> SEQ ID NO 366 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 366 tttggataac gacctgaaga 20<210> SEQ ID NO 367 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Homo sapiens <400> SEQUENCE: 367 ctcctgcaag gaggggacct 20 <210> SEQ IDNO 368 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens<400> SEQUENCE: 368 gtgttctatg agctggccca 20 <210> SEQ ID NO 369 <211>LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE:369 tggcagcacc tcacatttga 20 <210> SEQ ID NO 370 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 370 ccgaagctgaccagcagatg 20 <210> SEQ ID NO 371 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Homo sapiens <400> SEQUENCE: 371 ggaccagact gaatccctgt 20<210> SEQ ID NO 372 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 372 ccctacgggg ttaaggaacc 20 <210> SEQ ID NO373 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 373 cggggttaag gaacccaggt 20 <210> SEQ ID NO 374 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 374ttaaggaacc caggtgctcc 20 <210> SEQ ID NO 375 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 375 gcgacaatgacagctgacaa 20 <210> SEQ ID NO 376 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 376 aatgacagct gacaaggaga 20<210> SEQ ID NO 377 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 377 gcaaggagac ggaggtcttc 20 <210> SEQ ID NO378 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 378 acagtgtgag ctcccacctg 20 <210> SEQ ID NO 379 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 379atgcgcctgg ccatcagctt 20 <210> SEQ ID NO 380 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 380 atggataacttgtacctgaa 20 <210> SEQ ID NO 381 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 381 aacttgtacc tgaaagcctt 20<210> SEQ ID NO 382 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 382 aagccttgga gggtttcatt 20 <210> SEQ ID NO383 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 383 catcagcaag ttcatgggac 20 <210> SEQ ID NO 384 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 384ttcatgggac ttacccaggt 20 <210> SEQ ID NO 385 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 385 ttacccaggtagaactaaca 20 <210> SEQ ID NO 386 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 386 caggacacag catctttgac 20<210> SEQ ID NO 387 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 387 atctttgact tcactcatcc 20 <210> SEQ ID NO388 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 388 accgagcgtg acttcttcat 20 <210> SEQ ID NO 389 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 389gacttcttca tgaggatgaa 20 <210> SEQ ID NO 390 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 390 cttcatgaggatgaagtgca 20 <210> SEQ ID NO 391 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 391 caacagaggc cggactgtca 20<210> SEQ ID NO 392 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 392 tgcaccgggc aagtgagagt 20 <210> SEQ ID NO393 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 393 aagtgagagt ctacaacaac 20 <210> SEQ ID NO 394 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 394atcatcatgt gtgagccaat 20 <210> SEQ ID NO 395 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 395 gtgagccaatccagcaccca 20 <210> SEQ ID NO 396 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 396 ttcctgagcc gccacagcat 20<210> SEQ ID NO 397 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 397 gaactgattg gttaccaccc 20 <210> SEQ ID NO398 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 398 agtcaccaga acttgtgcac 20 <210> SEQ ID NO 399 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 399ccagaacttg tgcaccaagg 20 <210> SEQ ID NO 400 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 400 tggctggagacccaggggac 20 <210> SEQ ID NO 401 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 401 ggagacccag gggacggtca 20<210> SEQ ID NO 402 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 402 cccaggggac ggtcatctac 20 <210> SEQ ID NO403 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 403 gggacggtca tctacaaccc 20 <210> SEQ ID NO 404 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 404gacgtggtgt tctccatgga 20 <210> SEQ ID NO 405 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 405 ggtgttctccatggaccaga 20 <210> SEQ ID NO 406 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 406 tggccatgaa cagcatcttt 20<210> SEQ ID NO 407 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 407 gagcaactac ctgttcacca 20 <210> SEQ ID NO408 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 408 aaggaggagc ccgaggagct 20 <210> SEQ ID NO 409 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 409cctcagccta tggcaaggcc 20 <210> SEQ ID NO 410 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 410 gcctatggcaaggccatcct 20 <210> SEQ ID NO 411 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 411 agctgctcca cgcccagcag 20<210> SEQ ID NO 412 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 412 cagtacccag acggacttca 20 <210> SEQ ID NO413 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 413 aagtacccgc agcagttgga 20 <210> SEQ ID NO 414 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 414ctgggagcaa agggtccctg 20 <210> SEQ ID NO 415 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 415 tgctgtggccaggccagcac 20 <210> SEQ ID NO 416 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 416 ctcatgtctc catgttcaag 20<210> SEQ ID NO 417 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 417 catgttcaag atgaggtctg 20 <210> SEQ ID NO418 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 418 tcaagatgag gtctgcaaag 20 <210> SEQ ID NO 419 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 419gccctgtcca acaagctgaa 20 <210> SEQ ID NO 420 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 420 tccaggcaccagcagttcac 20 <210> SEQ ID NO 421 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 421 atgaagagcc tcatgggcgg 20<210> SEQ ID NO 422 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 422 tcatgggcgg gacctgtcct 20 <210> SEQ ID NO423 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 423 gacctgtcct ttgatgcctg 20 <210> SEQ ID NO 424 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 424aaggtgtcag gcgtggccag 20 <210> SEQ ID NO 425 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 425 agtcgactgctggggccatc 20 <210> SEQ ID NO 426 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 426 actgctgggg ccatcgttcg 20<210> SEQ ID NO 427 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 427 ccatcgttcg agccttacct 20 <210> SEQ ID NO428 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 428 gaactgacca gatatgactg 20 <210> SEQ ID NO 429 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 429gaccagatat gactgtgagg 20 <210> SEQ ID NO 430 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 430 gatatgactgtgaggtgaac 20 <210> SEQ ID NO 431 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 431 gactgtgagg tgaacgtgcc 20<210> SEQ ID NO 432 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 432 tgaggtgaac gtgcccgtgc 20 <210> SEQ ID NO433 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 433 acactcctgc aggggagaga 20 <210> SEQ ID NO 434 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 434aggggagaga ccttctcaga 20 <210> SEQ ID NO 435 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 435 ccttctcagagctctggacc 20 <210> SEQ ID NO 436 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 436 ctggaccagg ccacctgagc 20<210> SEQ ID NO 437 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Musmusculus <400> SEQUENCE: 437 aactcaagtg ttcttgttct 20 <210> SEQ ID NO438 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400>SEQUENCE: 438 actgtcatgt ctcaactgtt 20 <210> SEQ ID NO 439 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 439tcaagatgag gttagtgaca 20 <210> SEQ ID NO 440 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Mus musculus <400> SEQUENCE: 440 cccagaaaagtgactcctgt 20 <210> SEQ ID NO 441 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Mus musculus <400> SEQUENCE: 441 cagtgtcctg agactgtatg 20<210> SEQ ID NO 442 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 442 ttcgcggctg gacgattcag 20 <210> SEQ ID NO443 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 443 cctcatggtc gcagggatga 20 <210> SEQ ID NO 444 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 444tctcctcatg gtcgcaggga 20 <210> SEQ ID NO 445 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 445 tcatggtcacatggatgagt 20 <210> SEQ ID NO 446 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <400> SEQUENCE: 446 cctcatggtc acatggatga 20 <210>SEQ ID NO 447 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <400> SEQUENCE: 447 ctcatggtca catggatgag 20 <210> SEQ ID NO448 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400>SEQUENCE: 448 atttcctcat ggtcacatgg 20 <210> SEQ ID NO 449 <211> LENGTH:20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE: 449aaaccctcca aggctttcag 20 <210> SEQ ID NO 450 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 450 tcctcatggtcgcagggatg 20 <210> SEQ ID NO 451 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Synthetic Construct <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (12)..(12) <223> OTHER INFORMATION: n = inosine <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: (15)..(15) <223> OTHERINFORMATION: n = pseudouridine <400> SEQUENCE: 451 tcctcatggt cncanggatg20 <210> SEQ ID NO 452 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: SyntheticConstruct <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:(11)..(11) <223> OTHER INFORMATION: n = inosine <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: (14)..(14) <223> OTHERINFORMATION: n = pseudouridine <400> SEQUENCE: 452 cctcatggtc ncanggatga20 <210> SEQ ID NO 453 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: ControlOligonucleotide <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (1)..(20) <223> OTHER INFORMATION: equal mixture of the basesA, C, G and T <400> SEQUENCE: 453 nnnnnnnnnn nnnnnnnnnn 20 <210> SEQ IDNO 454 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Control Oligonucleotide<400> SEQUENCE: 454 ccttccctga aggttcctcc 20 <210> SEQ ID NO 455 <211>LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Synthetic Construct <400> SEQUENCE:455 cgagaggcgg acgggaccg 19 <210> SEQ ID NO 456 <211> LENGTH: 21 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (20)..(21) <223> OTHER INFORMATION:deoxythymidine <400> SEQUENCE: 456 cgagaggcgg acgggaccgt t 21 <210> SEQID NO 457 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic Construct<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (20)..(21)<223> OTHER INFORMATION: deoxythymidine <400> SEQUENCE: 457 cggtcccgtccgcctctcgt t 21 <210> SEQ ID NO 458 <211> LENGTH: 19 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Synthetic Construct <400> SEQUENCE: 458 cggtcccgtccgcctctcg 19

What is claimed is:
 1. A compound 8 to 80 nucleobases in length targetedto a nucleic acid molecule encoding HIF1α, wherein said compoundspecifically hybridizes with said nucleic acid molecule encoding HIF1αand inhibits the expression of HIF1α.
 2. The compound of claim 1comprising 12 to 50 nucleobases in length.
 3. The compound of claim 2comprising 15 to 30 nucleobases in length.
 4. The compound of claim 1comprising an oligonucleotide.
 5. The compound of claim 4 comprising anantisense oligonucleotide.
 6. The compound of claim 4 comprising a DNAoligonucleotide.
 7. The compound of claim 4 comprising an RNAoligonucleotide.
 8. The compound of claim 4 comprising a chimericoligonucleotide.
 9. The compound of claim 4 wherein at least a portionof said compound hybridizes with RNA to form an oligonucleotide-RNAduplex.
 10. The compound of claim 1 having at least 70% complementaritywith a nucleic acid molecule encoding HIF1α (SEQ ID NO: 4) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 11.The compound of claim 1 having at least 80% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 4) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 12.The compound of claim 1 having at least 90% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 4) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 13.The compound of claim 1 having at least 95% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 4) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 14.The compound of claim 1 having at least 70% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 133) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 15.The compound of claim 1 having at least 80% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 133) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 16.The compound of claim 1 having at least 90% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 133) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 17.The compound of claim 1 having at least 95% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 133) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 18.The compound of claim 1 having at least 70% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 206) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 19.The compound of claim 1 having at least 80% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 206) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 20.The compound of claim 1 having at least 90% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 206) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 21.The compound of claim 1 having at least 95% complementarity with anucleic acid molecule encoding HIF1α (SEQ ID NO: 206) said compoundspecifically hybridizing to and inhibiting the expression of HIF1α. 22.The compound of claim 1 having at least one modified internucleosidelinkage, sugar moiety, or nucleobase.
 23. The compound of claim 1 havingat least one 2′-O-methoxyethyl sugar moiety.
 24. The compound of claim 1having at least one phosphorothioate internucleoside linkage.
 25. Thecompound of claim 1 having at least one 5-methylcytosine.
 26. Thecompound of claim 1 comprising at least an 8-nucleobase portion of SEQID NO: SEQ ID NOs 14, 15, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28,29, 30, 31, 32, 35, 36, 40, 41, 42, 43, 44, 45, 47, 48, 49, 50, 51, 53,55, 57, 58, 59, 60, 61, 63, 64, 66, 67, 69, 70, 71, 74, 75, 78, 83, 134,135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148,149, 150, 151, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163,166, 167, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 192, 193, 194, 197,198, 200, 201, 202, 203 or
 207. 27. The compound of claim 1 having asequence selected from the group consisting of SEQ ID NO: SEQ ID NOs 14,15, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 29, 30, 31, 32, 35, 36,40, 41, 42, 43, 44, 45, 47, 48, 49, 50, 51, 53, 55, 57, 58, 59, 60, 61,63, 64, 66, 67, 69, 70, 71, 74, 75, 78, 83, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 153,154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 166, 167, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 192, 193, 194, 197, 198, 200, 201, 202,203 and
 207. 28. The compound of claim 1, wherein said compoundcomprises an antisense nucleic acid molecule that is specificallyhybridizable with a 5′-untranslated region (5′UTR) of hypoxia-induciblefactor 1 alpha (SEQ ID NO: 4).
 29. The compound of claim 1, wherein saidcompound comprises an antisense nucleic acid molecule that isspecifically hybridizable with a start region of hypoxia-induciblefactor 1 alpha (SEQ ID NO: 4).
 30. The compound of claim 1, wherein saidcompound comprises an antisense nucleic acid molecule that isspecifically hybridizable with a coding region of hypoxia-induciblefactor 1 alpha (SEQ ID NO: 4).
 31. The compound of claim 1, wherein saidcompound comprises an antisense nucleic acid molecule that isspecifically hybridizable with a stop region of hypoxia-inducible factor1 alpha (SEQ ID NO: 4).
 32. The compound of claim 1, wherein saidcompound comprises an antisense nucleic acid molecule that isspecifically hybridizable with a 3′-untranslated region ofhypoxia-inducible factor 1 alpha (SEQ ID NO: 4).
 33. A method ofinhibiting the expression of HIF1α in cells or tissues comprisingcontacting said cells or tissues with the compound of claim 1 so thatexpression of HIF1α is inhibited.
 34. A method of screening for amodulator of HIF1α, the method comprising the steps of: a. contacting apreferred target segment of a nucleic acid molecule encoding HIF1α withone or more candidate modulators of HIF1α, and b. identifying one ormore modulators of HIF1α expression which modulate the expression ofHIF1α.
 35. The method of claim 19 wherein the modulator of HIF1αexpression comprises an oligonucleotide, an antisense oligonucleotide, aDNA oligonucleotide, an RNA oligonucleotide, an RNA oligonucleotidehaving at least a portion of said RNA oligonucleotide capable ofhybridizing with RNA to form an oligonucleotide-RNA duplex, or achimeric oligonucleotide.
 36. A diagnostic method for identifying adisease state comprising identifying the presence of HIF1α in a sampleusing at least one of the primers comprising SEQ ID NOs 5 or 6, or theprobe comprising SEQ ID NO:
 7. 37. A kit or assay device comprising thecompound of claim
 1. 38. A method of treating an animal having a diseaseor condition associated with HIF1α comprising administering to saidanimal a therapeutically or prophylactically effective amount of thecompound of claim 1 so that expression of HIF1α is inhibited.
 39. Themethod of claim 38 wherein the disease or condition is ahyperproliferative disorder.
 40. The method of claim 39 wherein thehyperproliferative disorder is cancer.
 41. The method of claim 40wherein the cancer carries a p53 mutation.
 42. The method of claim 39wherein the hyperproliferative disorder is an angiogenic disorder. 43.The method of claim 42 wherein the angiogenic disorder affects the eye.44. A composition comprising the compound of claim 1 in apharmaceutically acceptable carrier.
 45. A compound of claim 1 whichalso inhibits the expression of HIF2α.
 46. A composition comprising acompound of claim 45 in a pharmaceutically acceptable carrier.
 47. Acompound 8 to 80 nucleobases in length targeted to a nucleic acidmolecule encoding HIF2α, wherein said compound specifically hybridizeswith said nucleic acid molecule encoding HIF2α and inhibits theexpression of HIF2α.
 48. The compound of claim 47 comprising 12 to 50nucleobases in length.
 49. The compound of claim 48 comprising 15 to 30nucleobases in length.
 50. The compound of claim 47 comprising anoligonucleotide.
 51. The compound of claim 50 comprising an antisenseoligonucleotide.
 52. The compound of claim 50 comprising a DNAoligonucleotide.
 53. The compound of claim 50 comprising an RNAoligonucleotide.
 54. The compound of claim 50 comprising a chimericoligonucleotide.
 55. The compound of claim 50 wherein at least a portionof said compound hybridizes with RNA to form an oligonucleotide-RNAduplex.
 56. The compound of claim 47 having at least 70% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 208) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 57. The compound of claim 47 having at least 80% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 208) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 58. The compound of claim 47 having at least 90% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 208) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 59. The compound of claim 47 having at least 95% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 208) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 60. The compound of claim 47 having at least 70% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 212) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 61. The compound of claim 47 having at least 80% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 212) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 62. The compound of claim 47 having at least 90% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 212) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 63. The compound of claim 47 having at least 95% complementaritywith a nucleic acid molecule encoding HIF2α (SEQ ID NO: 212) saidcompound specifically hybridizing to and inhibiting the expression ofHIF2α.
 64. The compound of claim 47 having at least one modifiedinternucleoside linkage, sugar moiety, or nucleobase.
 65. The compoundof claim 47 having at least one 2′-O-methoxyethyl sugar moiety.
 66. Thecompound of claim 47 having at least one phosphorothioateinternucleoside linkage.
 67. The compound of claim 47 having at leastone 5-methylcytosine.
 68. The compound of claim 47 comprising at leastan 8-nucleobase portion of SEQ ID NO: SEQ ID NOs 219, 220, 221, 211,223, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254 or
 255. 69. The compound of claim 47 having a sequence selectedfrom the group consisting of SEQ ID NO: SEQ ID NOs 219, 220, 221, 211,223, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254 and
 255. 70. The compound of claim 47 comprising at least an8-nucleobase portion of SEQ ID NO: 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 305, 306, 307, 308, 309, 310, 311,313, 314, 315, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327,328, 329, 331, 332, 333, 334 or
 335. 71. The compound of claim 47 havinga sequence selected from the group consisting of SEQ ID NO, 260, 261,262, 263, 264, 265, 266, 267, 268, 269, 270, 272, 273, 274, 275, 276,277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290,291, 292, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 305, 306,307, 308, 309, 310, 311, 313, 314, 315, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 331, 332, 333, 334 and
 335. 72. Thecompound of claim 47, wherein said compound comprises an antisensenucleic acid molecule that is specifically hybridizable with a5′-untranslated region (5′UTR) of hypoxia-inducible factor 2 alpha (SEQID NO: 208).
 73. The compound of claim 47, wherein said compoundcomprises an antisense nucleic acid molecule that is specificallyhybridizable with a start region of hypoxia-inducible factor 2 alpha(SEQ ID NO: 208).
 74. The compound of claim 47, wherein said compoundcomprises an antisense nucleic acid molecule that is specificallyhybridizable with a coding region of hypoxia-inducible factor 2 alpha(SEQ ID NO: 208).
 75. The compound of claim 47, wherein said compoundcomprises an antisense nucleic acid molecule that is specificallyhybridizable with a stop region of hypoxia-inducible factor 2 alpha (SEQID NO: 208).
 76. The compound of claim 47, wherein said compoundcomprises an antisense nucleic acid molecule that is specificallyhybridizable with a 3′-untranslated region of hypoxia-inducible factor 2alpha (SEQ ID NO: 208).
 77. A method of inhibiting the expression ofHIF2α in cells or tissues comprising contacting said cells or tissueswith the compound of claim 47 so that expression of HIF2α is inhibited.78. A method of screening for a modulator of HIF2α, the methodcomprising the steps of: a. contacting a preferred target segment of anucleic acid molecule encoding HIF2α with one or more candidatemodulators of HIF2α, and b. identifying one or more modulators of HIF2αexpression which modulate the expression of HIF2α.
 79. The method ofclaim 78 wherein the modulator of HIF2α expression comprises anoligonucleotide, an antisense oligonucleotide, a DNA oligonucleotide, anRNA oligonucleotide, an RNA oligonucleotide having at least a portion ofsaid RNA oligonucleotide capable of hybridizing with RNA to form anoligonucleotide-RNA duplex, or a chimeric oligonucleotide.
 80. Adiagnostic method for identifying a disease state comprising identifyingthe presence of HIF2α in a sample using at least one of the primerscomprising SEQ ID NOs 209 or 210, or the probe comprising SEQ ID NO:211.
 81. A kit or assay device comprising the compound of claim
 47. 82.A method of treating an animal having a disease or condition associatedwith HIF2α comprising administering to said animal a therapeutically orprophylactically effective amount of the compound of claim 47 so thatexpression of HIF2α is inhibited.
 83. The method of claim 82 wherein thedisease or condition is a hyperproliferative disorder.
 84. The method ofclaim 83 wherein the hyperproliferative disorder is cancer.
 85. Themethod of claim 84 wherein the cancer carries a p53 mutation.
 86. Themethod of claim 83 wherein the hyperproliferative disorder is anangiogenic disorder.
 87. The method of claim 86 wherein the angiogenicdisorder affects the eye.
 88. A composition comprising the compound ofclaim 47 in a pharmaceutically acceptable carrier.
 89. A compound ofclaim 47 which also inhibits the expression of HIF1α.
 90. A compositioncomprising a compound of claim 89 in a pharmaceutically acceptablecarrier.
 91. An antisense compound which inhibits the expression ofHIF1α and HIF2α.
 92. The antisense compound of claim 91 comprising SEQID NO: 443, 444, 233, 141, 445, 446, 447, 448, 449 or
 450. 93. Theantisense compound of claim 91 which comprises at least one universalbase.
 94. The antisense compound of claim 93 wherein the universal baseis inosine or 3-nitropyrrole.
 95. A composition comprising the compoundof claim 91 and a pharmaceutically acceptable carrier.
 96. A method ofinhibiting the expression of HIF1α and HIF2α in cells or tissuescomprising contacting said cells or tissues with the compound of claim91 so that expression of HIF1α and HIF2α is inhibited.
 97. A method ofinhibiting the expression of HIF1α and HIF2α in cells or tissuescomprising contacting said cells or tissues with (i) a first compound 8to 80 nucleobases in length targeted to a nucleic acid molecule encodingHIF1α, wherein said first compound specifically hybridizes with saidnucleic acid molecule encoding HIF1α and inhibits expression of HIF1α,and (ii) a second compound 8 to 80 nucleobases in length targeted to anucleic acid molecule encoding HIF2α, wherein said second compoundspecifically hybridizes with said nucleic acid molecule encoding HIF2αand inhibits expression of HIF2α, so that expression of HIF1α and HIF2αis inhibited.
 98. A method of modulating hypoxia-inducible geneexpression in cells or tissues comprising contacting said cells ortissues with (i) a first compound 8 to 80 nucleobases in length targetedto a nucleic acid molecule encoding HIF1α, wherein said first compoundspecifically hybridizes with said nucleic acid molecule encoding HIF1αand inhibits expression of HIF1α, and (ii) a second compound 8 to 80nucleobases in length targeted to a nucleic acid molecule encodingHIF2α, wherein said second compound specifically hybridizes with saidnucleic acid molecule encoding HIF2α and inhibits expression of HIF2α,so that expression of HIF1α and HIF2α is inhibited.
 99. The method ofclaim 98 wherein said cells or tissues are cancer cells or tissues. 100.The method of claim 99 wherein the cancer cells or tissues carry a p53mutation.
 101. A method of modulating hypoxia-inducible gene expressionin cells or tissues comprising contacting said cells or tissues with acompound of claim 91 so that expression of HIF1α and HIF2α is inhibited.102. The method of claim 101 wherein said cells or tissues are cancercells or tissues.
 103. The method of claim 102 wherein the cancer cellsor tissues carry a p53 mutation.
 104. A method of treating an animalhaving a disease or condition associated with hypoxia or ahypoxia-inducible factor or a hypoxia inducible gene comprisingadministering to said animal (i) a therapeutically or prophylacticallyeffective amount of a first compound 8 to 80 nucleobases in lengthtargeted to a nucleic acid molecule encoding HIF1α, wherein said firstcompound specifically hybridizes with said nucleic acid moleculeencoding HIF1α and inhibits expression of HIF1α, and (ii) atherapeutically or prophylactically effective amount of a secondcompound 8 to 80 nucleobases in length targeted to a nucleic acidmolecule encoding HIF2α, wherein said second compound specificallyhybridizes with said nucleic acid molecule encoding HIF2ααand inhibitsexpression of HIF2α, so that expression of HIF1α and HIF2α is inhibited.105. The method of claim 104 wherein the disease or condition is ahyperproliferative disorder.
 106. The method of claim 105 wherein thehyperproliferative disorder is cancer.
 107. The method of claim 106wherein the cancer carries a p53 mutation.
 108. The method of claim 105wherein the hyperproliferative disorder is an angiogenic disorder. 109.The method of claim 108 wherein the angiogenic disorder affects the eye.110. A method of treating an animal having a disease or conditionassociated with hypoxia or a hypoxia-inducible factor or a hypoxiainducible gene comprising administering to said animal a therapeuticallyor prophylactically effective amount of a compound of claim 91 so thatexpression of HIF1α and HIF2α is inhibited.
 111. The method of claim 110wherein the disease or condition is a hyperproliferative disorder. 112.The method of claim 111 wherein the hyperproliferative disorder iscancer.
 113. The method of claim 112 wherein the cancer carries a p53mutation.
 114. The method of claim 111 wherein the hyperproliferativedisorder is an angiogenic disorder.
 115. The method of claim 114 whereinthe angiogenic disorder affects the eye.